Halogen treatment of heart attack and ischemic injury

ABSTRACT

The present invention relates to the use of halogen compounds, including iodide, and chalcogenide compounds, including iodide, sulfide and selenide, to treat and prevent diseases and injuries. The present invention further relates to compositions comprising a halogen compound and/or a chalcogenide compound, including pharmaceutical compositions, as well as methods of manufacturing such compounds and administering such compositions to subjects in need thereof.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No.61/937,943, filed on Feb. 10, 2014, U.S. Provisional Application No.62/007,015, filed on Jun. 3, 2014, U.S. Provisional Application No.62/060,338, filed on Oct. 6, 2014, and U.S. Provisional Application No.62/082,957, filed on Nov. 21, 2014, each of which is incorporated byreference herein in its entirety.

GOVERNMENT INTEREST

This invention was made with government support under D12AP00008 awardedby the Department of the Interior. The government has certain rights inthe invention.

FIELD OF THE INVENTION

This invention relates to compositions comprising halogen and/orchalcogenide compounds, including those comprising a halogen and/or achalcogen compound in a reduced form, e.g. halides and/or chalcogenides,and methods for treating or preventing injuries and diseases, includingdiseases and injuries associated with hypoxia, ischemia or reperfusioninjury and/or the formation of reactive oxygen species, e.g., heartattack, chronic heart failure, diseases and injuries associated withexcessive metabolic rate, e.g., epilepsy, and diseases and injuriesassociated with an undesired immune or inflammatory response, e.g.,graft versus host disease (GVHD) or organ transplant. The presentinvention also relates to methods for reducing or inhibiting an immuneresponse, using a composition comprising a halide and/or a chalcogenidecompound.

BACKGROUND OF THE INVENTION

Compounds containing a halogen element, i.e., those in Group 17 of theperiodic table are commonly termed “halogens” or “halogen compounds.”These elements are iodine (I), fluorine (F), chlorine (Cl), bromine (Br)and astatine (At). The artificially created element 117 (ununseptium)may also be a halogen. The term “halogen” means “salt-former,” andcompounds containing halogens are usually called “salts.” All halogenshave 7 electrons in their outer shells, giving them an oxidation numberof −1. The halogens all form binary compounds with hydrogen known as thehydrogen halides: hydrogen fluoride (HF), hydrogen chloride (HCl),hydrogen bromide (HBr), hydrogen iodide (HI), and hydrogen astatide(HAt). When in aqueous solution, the hydrogen halides are known ashydrohalic acids. The names of these acids are as follows: hydrofluoricacid, hydrochloric acid, hydrobromic acid, and hydroiodic acid. Chlorineplays a biological role in higher animal life as aqueous chlorideanions, which are included in the electrolyte makeup of inter- andintra-cellular fluids. Iodine is known to be needed by our bodies intrace amounts, particularly by the thyroid gland, and iron deficiencycan cause goiter. The other three halogens have no known biologicalrole.

In medicine, ischemia-reperfusion injuries are commonly associated withblood loss or reduced blood flow to a tissue or organ.Ischemia-reperfusion injury is a complex phenomenon often encounteredwhen blood flow is restricted to an organ or tissue due to injury orblood loss, and also in surgical practice. Ischemia generally refers toa restriction in blood supply to tissues, causing a shortage of oxygenand glucose needed for cellular metabolism. Ischemia is generally causedby problems with blood vessels, with resultant damage to or dysfunctionof tissue. It also refers to local anemia in a tissue or organ, whichmay result from injury or congestion (e.g., vasoconstriction, thrombosisor embolism). The consequences of such injury are local and remotetissue destruction, and sometimes death. Reperfusion refers generally tothe restoration of blood flow to an ischemic tissue or organ. Primaryreperfusion therapies, including primary percutaneous coronaryintervention (PCI) and thrombolysis, are the standard of care for thetreatment of acute coronary syndromes. Prompt restoration of blood flowto ischemic myocardium limits infarct size and reduces mortality.Unfortunately, however, the return of blood flow can also result intissue, e.g., cardiac, damage and complications, referred to asreperfusion injury. Reperfusion injury is the tissue damage caused whenblood supply returns to the tissue after a period of ischemia or lack ofoxygen. The absence of oxygen and nutrients from blood during theischemic period creates a condition in which the restoration ofcirculation results in inflammation and oxidative damage through theinduction of oxidative stress rather than restoration of normalfunction. For example, in myocardial ischemia, considerable evidenceattributes reactive oxygen species (ROS), produced either by themyocardium itself or by infiltrating inflammatory cells, as an earlyevent in this process. Once produced, ROS can lead to cellular damagethrough a number of pathways including direct damage to membranes andproteins or indirect damage through the activation of pro-apoptoticpathways.

Effective therapies to reduce or prevent ischemic and reperfusioninjuries have proven elusive. Despite an improved understanding of thepathophysiology of these processes and encouraging preclinical trials ofmultiple agents, most of the clinical trials to preventischemia-reperfusion injury have been disappointing. Accordingly,therapies for treating or limiting injuries and damage resulting fromhypoxia, ischemia and/or reperfusion remain an active area ofinvestigation.

Clearly, there is a need in the art for new pharmaceutical compositionsand methods for treating or preventing ischemic and/or reperfusioninjuries, e.g., including those that may be conveniently administered topatients, both in a controlled medical environment e.g., for treatmentof disease, as a treatment in the field during an emergency, or incritical care in response to a catastrophic injury or life-threateningmedical event. The present invention meets this need by providingpharmaceutical compositions comprising active agents, which aredemonstrated herein to protect animals from injury resulting fromhypoxic and/or ischemic conditions, as well as other injuries anddisease conditions.

BRIEF SUMMARY OF THE INVENTION

The present invention provides compositions and methods useful intreating or preventing a variety of diseases, conditions, and disorders.

In one embodiment, the present invention provides a method for treatingor preventing an injury or disease in a subject, comprising providing tosaid subject a composition comprising a halogen compound and apharmaceutically acceptable carrier, diluent, or excipient. In certainembodiments, the halogen compound is in a chemically reduced form. Incertain embodiments, the halogen compound comprises iodine, bromine,chlorine, fluorine, or astatine. In particular embodiments wherein thehalogen compound comprises iodine, said halogen compound is an iodide.In various embodiments, the iodide is sodium iodide, potassium iodide,hydrogen iodide, calcium iodide, or silver iodide. In particularembodiments, the halogen compound is an iodate. In various embodiments,the iodate is sodium iodate, potassium iodate, calcium iodate, or silveriodate. In certain embodiments, the halogen compound comprises bromine.In particular embodiments wherein the halogen compound comprises abromine, the halogen compound is a bromide. In certain embodiments, thehalogen is present in the halogen compound in a reduced form.

In certain embodiments of the present invention, the compositioncomprises one or more reducing agents or antioxidants. In particularembodiments, the composition is formulated to maintain the halogen orhalogen compound present in the composition in a reduced state, e.g., tomaintain iodide in its −1 oxidation state. In one embodiment, thecomposition is a pharmaceutical composition, comprising a reduced formof a halogen compound, e.g., a reduced form of iodine, such as iodide,glutathione, and a pharmaceutically acceptable carrier, diluent, orexcipient. In certain embodiments, the glutathione is present in anamount sufficient to maintain the halogen or halogen compound in itsreduced state. In certain embodiments of methods and composition of thepresent invention, at least 90% of the reduced form of halogen orhalogen compound in the composition remains in a reduced form for atleast one hour, at least one week, at least one month, or at least sixmonths when stored at room temperature. In certain embodiments, at least90% of the reduced form of halogen or halogen compound in thecomposition is present in a reduced form for at least one month, atleast two months, at least four months, at least six months, or at leastone year when stored at about 4° C. In particular embodiments, at least50%, at least 75%, or at least 90% of the glutathione initially presentin the composition is in a reduced form.

In various embodiments of methods of the present invention, thecomposition is provided to the subject parenterally or orally. Incertain embodiments, the composition is provided to the subjectintravenously or by infusion.

In certain embodiments, the composition is formulated for oraladministration, and the composition comprises a stable reduced form ofthe halogen compound.

In particular embodiments, the composition is formulated for intravenousadministration or administration by infusion, and the compositioncomprises a stable reduced form of the halogen compound.

In various embodiments, the composition is provided to the subject in anamount sufficient to increase the blood concentration of the halogen atleast 100%, at least 200%, at least 300%, at least 400%, or at least500% for at least some time.

In various embodiments, the disease or injury is selected from any ofthe following, or is caused by or results from any of the following:ablation therapy, adrenalectomy, aortic aneurysm, aortic root surgery,aortic stenosis, aortic valve disease, arrhythmia, atherosclerosis,atrial flutter, atrial fibrillation, atrial septal defect, arteriovenousmalformation, awake brain surgery, bariatric surgery, bone marrowtransplant, brachial plexus injuries, bradycardia, brain aneuryism,breast augmentation surgery, breast reduction surgery, burn injury,coronary bypass surgery, coronary artery disease, cardiac ablation,cardiac catheterization, cardiac resynchronization therapy, cardiacsurgery, cardiomyopathy, cardiac surgery, cardiovascular diseases,carotid angioplasty and stenting, coarctation of the aorta, congenitalheart disease, coronary bypass surgery, coronary artery disease,critical care medicine, chronic obstructive pulmonary disease, elbowreplacement surgery, emergency medicine, general internal medicine,general surgery, gastrointestinal bleeding, heart attack, hearttransplant, heart valve surgery, hip replacement surgery, hypertrophiccardiomyopathy, hypoxia ischemia encephalopathy, hysterectomy, ileoanalanastomosis (j pouch) surgery, inflammatory bowel disease, ischemicheart disease, ischemia reperfusion injury, irritable bowel syndrome,jaw surgery, kidney transplant, laryngotracheal reconstruction surgery,liver transplant, lung volume reduction surgery, lung transplant,minimally invasive heart surgery, neurosurgery, oral and maxillofacialsurgery, orthopedic surgery, pancreas transplant, pancreatitis, partialnephrectomy, pediatric cervical spine surgery, pediatric surgery, pelvicorgan prolapse, plastic and reconstructive surgery, pulmonary andcritical care medicine, pulmonary atresia, pulmonary vein isolation,rectal prolapse, restrictive cardiomyopathy, retinal detachment,retinopathy of prematurity, robotic surgery, spinal cord injury,spontaneous coronary artery dissection, spontaneous occlusion of thecircle of willis, stroke, stroke telemedicine (telestroke), suddencardiac arrest, stereotactic radiosurgery, surgery, systolic heartfailure, chronic heart failure, tachycardia, teare's disease, thoracicaortic aneurysm, thoracic surgery, total elbow arthroplasty, tricuspidvalve disease, ulcerative colitis, valve-preserving aortic root repair,vascular and endovascular surgery, vascular medicine, or ventriculartachycardia. In certain embodiments, the injury or disease is selectedfrom, results from or is caused by inflammation, heart attack, coronarybypass surgery, ischemia, gut ischemia, liver ischemia, kidney ischemia,hypoxic-ischemic encephalopathy, stroke, traumatic brain injury, limbischemia, eye ischemia, sepsis, smoke, burn, reperfusion, or acute lunginjury. In certain embodiments, the injury is an infarct caused by aheart attack or a stroke. In certain embodiments, the disease ishypoxic-ischemic encephalopathy. In particular embodiments, the injuryis a medical procedure, such as e.g., a cell, tissue or organtransplantation, or results from a medical procedure, such as, e.g.,transplant rejection. In certain embodiments, the injury is anautologous or heterologous transplantation, e.g., of cells, tissue or anorgan. In certain embodiments, the injury is a coronary bypass surgery,optionally a coronary artery bypass graft (CABG).

Certain embodiments of methods of the present invention compriseproviding to the subject a composition comprising a halogen compound anda composition comprising one or more additional active agent. In certainembodiments, a single composition comprises both the halogen compoundand the one or more additional active agent, whereas in otherembodiments, the halogen and the one or more additional active agent arepresent in separate compositions. In particular embodiments,compositions of the present invention comprise a halogen compound and/orone or more additional active agents, and a pharmaceutically acceptablecarrier, diluent, or excipient. In particular embodiments, compositionsof the present invention are pharmaceutical compositions. In particularembodiments, the invention includes a composition comprising a halogencompound, optionally in a reduced form, one or more additional activeagent, and a pharmaceutically acceptable carrier, diluent, or excipient.In particular embodiments, the one or more additional active agentcomprises a chalcogenide, optionally in a reduced form. In particularembodiments, the chalcogenide comprises sulfide or selenide. Inparticular embodiments, the sulfide or selenide is a reduced form ofsulfide or selenide. In some embodiments, the reduced form of sulfide ispresent in a stable liquid pharmaceutical composition comprising apharmaceutically acceptable carrier, diluent, or excipient. In certainembodiments, the composition comprises both a halogen compound and achalcogenide, wherein the composition is formulated for intravenous ororal administration.

In particular embodiments of methods of the present invention, thecomposition comprises a carrier. In certain embodiments, the halogencompound is associated with the carrier. In certain embodiments wherethe composition comprises a halogen compound and one or more additionalactive agents, the halogen compound and the one or more additionalactive agent are associated with the carrier. In certain embodiments,the halogen compound and the one or more additional active agent presentin the composition are associated with the carrier covalently ornon-covalently. In particular embodiments, the carrier is albumin,plasma, serum, alpha-2-macroglobulin, or immunoglobulin, or apolypeptide related to any of these polypeptides.

In a related embodiment, the present invention provides a compositioncomprising a halogen compound and a carrier, wherein said halogencompound is associated to the carrier. In certain embodiments, thecomposition further comprises one or more additional active agents. Inparticular embodiments, the one or more additional active agents areassociated with the carrier. In certain embodiments, said halogencompound and/or said one or more additional active agent are associatedwith the carrier covalently or non-covalently. In certain embodiments,the one or more additional active agent comprises a chalcogenide. Inparticular embodiments, the chalcogenide comprises sulfide or selenide.In further embodiments, the sulfide or selenide is a reduced form ofsulfide or selenide.

In another related embodiments, the present invention includes acomposition comprising a stable reduced form of a halogen compound,wherein said pharmaceutical composition is formulated for intravenousadministration, administration by infusion, or oral administration. Inparticular embodiments, the halogen compound comprises iodine, bromine,chlorine, fluorine, or astatine. In certain embodiments, the halogencompound comprises iodine. In particular embodiments, the halogencompound is an iodide, an iodate, an organoiodide, a periodate, or aperiodinane. In certain embodiments, the iodide is sodium iodide,potassium iodide, hydrogen iodide, calcium iodide, zinc iodide, orsilver iodide. In certain embodiments, the iodate is sodium iodate,potassium iodate, calcium iodate, or silver iodate. In particularembodiments, the halogen compound comprises bromine. In certainembodiments, the halogen compound is a bromide. In particularembodiments, the composition comprises glutathione.

In certain embodiments of methods and composition of the presentinvention, at least 90% of the halogen compound in the composition ispresent in a reduced form for at least one hour, at least one week, atleast one month, or at least six months when stored at room temperature.In certain embodiments, at least 90% of the halogen compound in thecomposition is present in a reduced form for at least one month, atleast two months, at least four months, at least six months, or at leastone year when stored at about 4° C.

In certain embodiments of methods and compositions of the presentinvention, the composition comprising the halogen compound and/or thecomposition comprising the additional active agent comprises one or moreof a reducing agent, a tonicity agent, a stabilizer, a surfactant, alycoprotectant, a polyol, an antioxidant, or a preservative.

In related embodiments, the present invention includes a unit dosageform of a composition of the invention, wherein said unit dosage form isformulated for oral administration. In particular embodiments, the unitdosage form is a pill, tablet, caplet or capsule. In certainembodiments, the unit dosage form comprises less than or equal to 150mg, less than or equal to 125 mg, less than or equal to 100 mg, lessthan or equal to 75 mg, less than or equal to 50 mg, less than or equalto 25 mg, or less than or equal to 10 mg of the halogen compound. Incertain embodiments, the unit dosage form comprises between about 1 mgand about 150 mg (including any interval in this range), between about 1mg and about 125 mg, between about 1 mg and about 100 mg, between about1 mg and about 75 mg, between about 1 mg and about 50 mg, between about1 mg and about 25 mg or between about 1 mg and about 10 mg of thehalogen compound. In certain embodiments, the unit dosage form comprisesabout 150 mg, about 125 mg, about 100 mg, about 75 mg, about 50 mg,about 25 mg or about about 10 mg of the halogen compound. In certainembodiments, the unit dosage form comprises less than or equal to 1000mg, less than or equal to 800 mg, less than or equal to 700 mg, lessthan or equal to 500 mg, less than or equal to 250 mg, less than orequal to 200 mg, or less than or equal to 150 mg of the halogencompound. In certain embodiments, the unit dosage form comprises betweenabout 100 mg and about 1000 mg (including any interval in this range),between about 150 mg and about 800 mg, between about 200 mg and about700 mg, between about 250 mg and about 600 mg, between about 300 mg andabout 500 mg, between about 350 mg and about 450 mg or between about 300mg and about 700 mg of the halogen compound. In certain embodiments, theunit dosage form comprises about 200 mg, about 300 mg, about 400 mg,about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, orabout about 1000 mg of the halogen compound. In certain embodiments, theunit dosage form comprises or contains an amount of halogen compoundand/or chalcogen compound to achieve an effective dose when one or moreof the unit dosage form is provided to a subject, e.g., a mammal, suchas a human. For example, one or more, two or more, or four or more, unitdosage forms may be provided to the subject in one day to achieve aneffective amount. In particular embodiments, the halogen compoundcomprises iodine, bromine, chlorine, fluorine, or astatine. In certainembodiments, the halogen compound comprises iodine. In certainembodiments, the halogen compound is an iodide, an iodate, anorganoiodide, a periodate, or a periodinane. In certain embodiments, thehalogen compound is an iodide. In certain embodiments, the iodide issodium iodide, potassium iodide, hydrogen iodide, calcium iodide, orsilver iodide. In certain embodiments, the halogen compound is aniodate. In certain embodiments, the iodate is sodium iodate, potassiumiodate, calcium iodate, or silver iodate. In certain embodiments, thehalogen compound comprises bromine. In certain embodiments, the halogencompound is a bromide.

In particular embodiments, the unit dosage form further comprises one ormore additional active agent. In certain embodiments, the one or moreadditional active agent comprises a chalcogenide. In certainembodiments, the chalcogenide comprises sulfide or selenide. In certainembodiments, the sulfide or selenide is a reduced form of sulfide orselenide.

The present invention further includes a method of treating orpreventing an injury or disease in a subject, comprising providing tosaid subject any of the unit dosage forms of the present invention. Inparticular embodiments, the disease or injury is any of those describedherein. In certain embodiments, the injury is an infarct caused by aheart attack or a stroke. In certain embodiments, the disease is chronicheart failure, e.g., systolic heart failure. In certain embodiments, theinjury is caused by coronary bypass surgery, optionally a coronaryartery bypass graft (CABG).

In certain embodiments of any of the methods of the present invention,the method further comprises providing to the subject a compositioncomprising a goitrogen, a compound that inhibits or impedes thyroidhormone production or activity, or glutathione. In particularembodiment, the one or more additional active agent comprisesglutathione. In certain embodiments, at least 50%, at least 75%, or atleast 90% of the glutathione provided is in a reduced state.

In certain embodiments, any of the compositions of the present inventionfurther comprises one or more additional active agents. In certainembodiments, the one or more additional active agent comprises agoitrogen, a compound that inhibits or impedes thyroid hormoneproduction or activity, or glutathione. In particular embodiment, theone or more additional active agent comprises glutathione. In certainembodiments, at least 50%, at least 75%, or at least 90% of theglutathione provided is in a reduced state.

In particular embodiments of the methods and compositions of theinvention, the goitrogen, compound that inhibits or impedes thyroidhormone production or activity, or glutathione is associated with thecarrier.

In certain embodiments, the present invention includes a method fortreating or preventing an injury or disease in a subject, comprisingproviding to said subject a composition comprising a chalcogenide andglutathione, or comprising a halogen compound and glutathione, whereinsaid composition further comprises a pharmaceutically acceptablecarrier, diluent, or excipient. In certain embodiments, the compositionfurther comprises a halogen compound or a chalcogenide, respectively. Incertain embodiments, the composition comprises a halogen compound, achalcogenide, and glutathione. In certain embodiments, the halogencompound comprises iodine, e.g., iodide or iodate. In certainembodiments, the chalcogenide comprises selenide or sulfide, includinge.g., reduced forms of either or both sulfide and selenide. In certainembodiments, the disease or injury is any of those described herein. Inparticular embodiments, the injury or disease is selected from, resultsfrom or is caused by inflammation, heart attack, chronic heart failure,coronary bypass surgery, ischemia, gut ischemia, liver ischemia, kidneyischemia, hypoxic-ischemic encephalopathy, stroke, traumatic braininjury, limb ischemia, eye ischemia, sepsis, smoke, burn, reperfusion,or acute lung injury. In one embodiment, the injury is an infarct causedby a heart attack or a stroke. In one embodiment, the injury is causedby coronary bypass surgery, optionally a coronary artery bypass graft(CABG).

In various embodiments of any of the methods of the invention, about 1pg/kg to about 1 g/kg of chalcogenide, sulfide, and/or selenide isprovided to the subject. In certain embodiments, about 10 μg/kg to about10 mg/kg of chalcogenide, e.g., sulfide or selenide, is provided to thesubject. In certain embodiments of any of the methods of the invention,the halogen compound is iodide, iodate, or iodine, and about 10 pg/kg toabout 1 g/kg of iodide, iodate, or iodine is provided to the subject. Inparticular embodiments, about 10 μg/kg to about 10 mg/kg, about 100μg/kg to about 10 mg/kg, about 1 mg/kg to about 10 mg/kg, about 1 mg/kgto about 5 mg/kg, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4mg/kg, about 5 mg/kg, about 6 mg/kg, about 7 mg/kg, about 8 mg/kg, about9 mg/kg, or about 10 mg/kg of iodide, iodate, or iodine is provided tothe subject. In certain embodiments, any of these amounts are providedto the subject within about 24 hours or within about 48 hours, e.g.,daily or every other day, for a duration of time, e.g., for one day, twodays, three days, four days, five days, six days, one week, two weeks,one month, two months, six months, one year, or longer.

In certain embodiments, the amount of halogen compound, e.g., iodide,iodate, or iodine, provided to the subject, e.g., a mammal or human, isless than or equal to 150 mg, less than or equal to 125 mg, less than orequal to 100 mg, less than or equal to 75 mg, less than or equal to 50mg, less than or equal to 25 mg, or less than or equal to 10 mg of thehalogen compound. In certain embodiments, the amount of halogen compoundprovided to the subject comprises between about 1 mg and about 150 mg(including any interval in this range), between about 1 mg and about 125mg, between about 1 mg and about 100 mg, between about 1 mg and about 75mg, between about 1 mg and about 50 mg, between about 1 mg and about 25mg or between about 1 mg and about 10 mg of the halogen compound. Incertain embodiments, the amount of halogen compound provided to thesubject is about 150 mg, about 125 mg, about 100 mg, about 75 mg, about50 mg, about 25 mg or about 10 mg of the halogen compound. In certainembodiments, the amount of halogen compound provided to the subject isless than or equal to 1000 mg, less than or equal to 800 mg, less thanor equal to 700 mg, less than or equal to 500 mg, less than or equal to250 mg, less than or equal to 200 mg, or less than or equal to 150 mg ofthe halogen compound. In certain embodiments, the amount of halogencompound provided to the subject comprises between about 100 mg andabout 1000 mg (including any interval in this range), between about 150mg and about 800 mg, between about 200 mg and about 700 mg, betweenabout 250 mg and about 600 mg, between about 300 mg and about 500 mg,between about 350 mg and about 450 mg or between about 300 mg and about700 mg of the halogen compound. In certain embodiments, the amount ofhalogen compound provided to the subject is about 200 mg, about 300 mg,about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg,about 900 mg, or about about 1000 mg of the halogen compound. In certainembodiments, any of these amounts are provided to the subject withinabout 24 hours or about 48 hours, e.g., daily or every other day, for aduration of time, e.g., for one day, two days, three days, four days,five days, six days, one week, two weeks, one month, two months, sixmonths, one year, or longer.

In various embodiments of any of the methods and composition of theinvention, the composition comprises about 1.5 μM to about 500 mMglutathione. In particular embodiments, the composition comprises about15 mM to about 500 mM glutathione.

In a further embodiment, the present invention comprises a compositioncomprising a chalcogenide and glutathione, or a halogen compound andglutathione, wherein said composition further comprises apharmaceutically acceptable carrier, diluent, or excipient. Inparticular embodiments, the composition further comprises a halogencompound or a chalcogenide, respectively. In certain embodiments, thecomposition comprises a halogen compound, a chalcogenide, andglutathione. In particular embodiments, the halogen compound comprisesiodine, e.g., iodide. In particular embodiments, the chalcogenidecomprises selenide or sulfide, e.g., a reduced form of either or bothselenide and sulfide. In certain embodiments, glutathione is present inthe composition at about 1 mM to about 500 mM. In certain embodiments,the composition is stored in a reduced oxygen or oxygen-free environmentprior to and/or while being provided to the subject. In certainembodiments, the composition is stored in said environment under argonor nitrogen prior to being provided to the subject. In certainembodiments, the composition is formulated for parenteral or oraladministration. In certain embodiments, the composition is formulatedfor intravenous administration or administration by infusion.

In another related embodiment, the present invention includes a methodfor treating or preventing a disease or injury in a subject, comprisinginhibiting or preventing a decrease in the ratio of the amount ofreduced glutathione to the amount of oxidized glutathione in thesubject's bloodstream or at the site of disease or injury. In certainembodiments, the method comprises increasing the ratio of the amount ofreduced glutathione to the amount of oxidized glutathione in thesubject's bloodstream or at the site of disease or injury. In particularembodiments, the method comprises providing a halogen compound and/or achalcogenide systemically to the subject prior to, at the time of, orafter onset of said disease or injury. In certain embodiments, thehalogen compound comprises iodine. In particular embodiments, thehalogen compound is an iodide or iodate. In certain embodiments, thechalcogenide comprises sulfur or selenium. In particular embodiments,the chalcogenide comprises selenide. In certain embodiments, the halogencompound and/or the chalcogenide is delivered to the subject orally orintravenously or by infusion. In certain embodiments, the halogencompound and/or the chalcogenide are delivered to the subject at a siteremote from said disease or injury. In certain embodiments, the diseaseor injury is localized within a certain tissue or organ. In certainembodiments, the disease or injury is localized within a cell type,tissue or organ. In certain embodiments, the method is used to treat orprevent sickle cell crisis or sickle cell anemia. In certainembodiments, the method is used to treat or prevent blood attack. Incertain embodiments, the method is used to treat or prevent myocardialinfarction or cardiogenic shock. In certain embodiments, the method isused to treat or prevent heart attack or chronic heart failure. Incertain embodiments, the method is used to treat or preventcontrast-induced nephropathy.

In a further embodiment, the present invention provides a method ofpreventing, inhibiting, or reducing an immune response in a subject,comprising providing a halogen compound and/or a chalcogenide to thesubject. In particular embodiments, the halogen compound comprisesiodine. In certain embodiments, the halogen compound is an iodide. Incertain embodiments, the chalcogenide comprises sulfur or selenium. Incertain embodiments, chalcogenide comprises selenide. In particularembodiments, the halogen compound and/or the chalcogenide is deliveredto the subject orally or intravenously or by infusion. In certainembodiments, the halogen compound and/or the chalcogenide are deliveredto the subject at a site of inflammation or immune reaction. In certainembodiments, the method is used to treat or prevent graft versus hostdisease. In certain embodiments, the method is used to inhibit or reducean immune response during or following cell, tissue or organtransplantation. In particular embodiments of any methods related tocell, tissue or organ transplantation, the cell, tissue or organ isallogeneic or autologous to the transplant recipient. In certainembodiments, the method is used to treat or prevent ischemic orreperfusion injury. In certain embodiments, the method reduces theamount of one or more reactive oxygen species in the subject'sbloodstream or at a site of inflammation or immune reaction within thesubject.

In certain embodiments, a goitrogen is used instead of or in addition toa chalcogenide or a halogen compound in the compositions and methods ofthe invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 provides a graph and table demonstrating NaI protection againstinfarct following ligation-induced ischemia. Ischemic conditions wereinduced in mice via open chest ligation of the left descending coronaryartery. In the graph, the Inf/AAR and Inf/LV bars represent morphometricanalysis of the infarct size (Inf) in relation to the total size of thearea at risk (AAR) or left ventricle (LV), respectively, in the presenceof saline control or increasing amounts of NaI (as indicated). TheAAR/LV bars shows the ratio of the size of the area at risk to the sizeof the left ventricle in the presence of saline control or increasingamounts of NaI (as indicated), and demonstrates that similar samplingwas performed on all test subjects. For each of the sets of bars forInf/AAR, Inf/LV and AAR/LV, the bars from left to right correspond tothe bars from top to bottom shown in the legend. The table beneath thegraph provides the mean volume (%) and standard error for each datapoint.

FIG. 2 provides a graph and table demonstrating NaI protection againstinfarct following ligation-induced ischemia. Ischemic conditions wereinduced in mice via open chest ligation of the left descending coronaryartery. In the graph, the Inf/AAR and Inf/LV bars represent morphometricanalysis of the infarct size (Inf) in relation to the total size of thearea at risk (AAR) or left ventricle (LV), respectively, in the presenceof saline control or increasing amounts of NaI (as indicated). TheAAR/LV bars shows the ratio of the size of the area at risk to the sizeof the left ventricle in the presence of saline control or increasingamounts of NaI (mg/kg), and demonstrates that similar sampling wasperformed on all test subjects. For each of the sets of bars forInf/AAR, Inf/LV and AAR/LV, the bars from left to right correspond tothe bars from top to bottom shown in the legend. The table beneath thegraph provides the mean volume (%) and standard error for each datapoint.

FIG. 3 provides a graph demonstrating protection from infarct byprophylactic treatment with NaI 5 minutes or 48 hours prior toligation-induced ischemia. Subjects were injected with either saline orNaI (1 mg/kg) 5 minutes or 48 hours prior to the induction of ischemiaby open chest ligation of the left descending coronary artery. After 60min of ischemic conditions, ligatures were released to inducereperfusion, and infarct size was measured and reported in relation tothe total size of the area at risk (AAR) and left ventricle (LV). Foreach of the sets of bars for Inf/AAR, Inf/LV and AAR/LV, the bars fromleft to right correspond to the bars from top to bottom shown in thelegend. The table beneath the graph provides the mean volume (%) andstandard error for each data point.

FIG. 4 provides a graph demonstrating protection from infarct byprophylactic treatment with NaI by a single intravenous administrationtwo days prior to ligation-induced ischemia as determined byechocardiogram. The graph shows the mean volume (%) for ejectionfraction and fractional shortening in normal animals (no infarctinduced), and animals treated with saline as control or with NaI. Foreach of the sets of bars for Ejection Fraction and FractionalShortening, the bars from left to right correspond to the bars from topto bottom shown in the legend.

FIG. 5 provides photographs demonstrating that glutathione preventsselenide from oxidation. The photographs show samples of 50 mM selenidein either water (FIG. 5A) or 150 mM GSH (FIG. 5B) over the eight minutesimmediately following preparation. The oxidized forms of selenide appeardark in the solutions, with the samples in glutathione clearly showingreduced levels of oxidation at each time point.

FIG. 6 provides a description of the experimental protocol demonstratingthe immunosuppressant effects of selenide and iodide.

FIG. 7 provides a table demonstrating the immunosuppressant propertiesof iodide and selenide.

FIG. 8 provides a graph and table demonstrating NaI protection againstinfarct following ligation-induced ischemia. In the graph, the Inf/AARand Inf/LV bars represent morphometric analysis of the infarct size(Inf) in relation to the total size of the area at risk (AAR) or leftventricle (LV), respectively, following ligation-induced ischemiainitiated after two days of a diet including drinking water containingthe indicated amounts of NaI. The AAR/LV bars shows the ratio of thesize of the area at risk to the size of the left ventricle afterdrinking normal water or water containing increasing amounts of NaI (asindicated), and demonstrates that similar sampling was performed on alltest subjects. For each of the sets of bars for Inf/AAR, Inf/LV andAAR/LV, the bars from left to right correspond to the bars from top tobottom shown in the legend. The amount of iodide present in the variousdrinking waters is indicated below the graph.

FIG. 9 provides graphs demonstrating NaI protection from chronic heartfailure following ligation-induced myocardial infarct. FIG. 9A is agraph showing left ventricle (LV) ejection fraction (%) at baseline andthe indicated times following induced myocardial infarct. A dietincluding drinking water containing 0.77 mg/kg of NaI was initiated justafter the ligation procedure was completed. FIG. 9B is a graph showingLV fractional shortening (%) at baseline and the indicated timesfollowing induced myocardial infarct. A diet including drinking watercontaining 0.77 mg/kg of NaI was initiated just after the ligationprocedure was completed. For each pair of bars, asterisks indicate astatistically significant difference between mice that received iodidecontaining (light bars) and normal drinking water (dark bars) (p<0.05).

FIG. 10 provides graphs demonstrating NaI protection from chronic heartfailure following ligation-induced myocardial infarct. FIG. 10A is agraph showing left ventricle (LV) ejection fraction (EF) (%) at baselineand the indicated times following induced myocardial infarct. A dietincluding drinking water containing 0.77 mg/kg of NaI was initiated justafter the ligation procedure was completed. FIG. 10B is a graph showingLV fractional shortening (FS) (%) at baseline and the indicated timesfollowing induced myocardial infarct. A diet including drinking watercontaining 0.77 mg/kg of NaI was initiated just after the ligationprocedure was completed. For each pair of bars, asterisks indicate astatistically significant difference between mice that received iodidecontaining (light bars) and normal drinking water (dark bars) (p<0.05).

FIG. 11 provides graphs showing plasma cardiac troponin I (cTpnI) levelsin the blood of animals treated with placebo, ischemic preconditioning,or 1 mg/kg NaI in a rat model of acute myocardial infarct. FIG. 11A is agraph showing cTpnI levels at 4 hours; FIG. 11B is a graph showing cTpnIlevels at 24 hours; and FIG. 11C is a graph showing cTPNI levels at both4 hours and 24 hours. *p<0.05 vs placebo by One Way ANOVA.

FIG. 12 provides graphs demonstrating successful NaI treatment of acutemyocardial infarct. In the graphs, the Inf/AAR and Inf/LV bars representmorphometric analysis of the infarct size (Inf) in relation to the totalsize of the area at risk (AAR) or the left ventricle (LV), respectively.FIG. 12A shows AAR size in animals treated with placebo, ischemicpreconditioning, or 1 mg/kg NaI; FIG. 12B shows % Inf/AAR in animalstreated with placebo, ischemic preconditioning, or 1 mg/kg NaI; and FIG.12C shows % Inf/LV in animals treated with placebo, ischemicpreconditioning, or 1 mg/kg NaI. *p<0.05 vs placebo by One Way ANOVA;and #p<0.05 vs ischemic preconditioning by One Way ANOVA.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based, in part, on the surprising andunexpected finding that halogen compounds, such as iodides, may be usedto treat or prevent diseases or injuries, including, e.g., thoseassociated with hypoxia, ischemia or reperfusion, formation of reactiveoxygen species, immune or inflammatory responses, or increased metabolicrate, including, e.g., chronic heart failure. As described in theaccompanying Examples, treatment with a halogen compound before, during,or after ischemic assault, such as heart attack, protects againstresulting tissue damage. In addition, the accompanying Examplesdemonstrate that treatment with a halogen improves the symptoms ofchronic heart failure, e.g., following a heart attack. Accordingly, thepresent invention provides new methods of treating and preventing avariety of diseases, conditions, and injuries, including thoseassociated with or resulting from hypoxia, ischemia and/or reperfusion,which comprise providing to a subject at risk of such disease,condition, or injury a composition comprising a halogen compound, suchas an iodide. In certain embodiments, the composition may be provided tothe subject prior to, concurrent with, or following the disease,condition, or injury. In addition, the present invention provides newcompositions (e.g., pharmaceutical compositions) comprising a halogencompound. In certain embodiments, the composition comprises the halogencompound in a stable, reduced form, such as, e.g., an iodide. Inparticular embodiments, the composition is formulated for intravenousadministration, administration by infusion, or oral administration. Themethods of the present invention are particularly advantageous, becausemany halogen compounds are known to be safe for use in mammals, so thepresent invention provides new and safe ways of treating and preventinga number of diseases, conditions and injuries.

In addition, the present invention includes compositions, e.g.,pharmaceutical compositions, comprising a stable, reduced form of ahalogen compound, e.g., an iodide. These compositions may be formulatedto maintain a halogen compound in a reduced form suitable foradministration to a subject and, thus, constitute new and improvedpharmaceutical products for treating and preventing a host of diseases,conditions and injuries, which have a shelf-life suitable fordistribution and storage prior to use. In addition to a halogencompound, compositions of the present invention may further comprise oneor more additional active agent, such as, e.g., a chalcogenide, whichmay also be in a stable, reduced form. Pharmaceutical compositions ofthe invention may be formulated for intravenous administration,infusion, or oral administration, in particular embodiments.

The present invention also includes unit dosage forms of a compositioncomprising an effective amount of a halogen compound, which are useful,inter alia, in treating a variety of diseases, conditions, and injuries.In various embodiments, the composition may be formulated forintravenous administration, administration by infusion, oraladministration and/or formulated to maintain the halogen compound in areduced form during storage. In particular embodiments, the unit dosageforms comprise or constitute pre-measured effective amounts of a halogencompound, which are advantageous for delivering an appropriate effectiveamount of a halogen compound to a subject, particularly during emergencyintervention at site of injury or during transfer of a subject. Incertain embodiments, the composition is a liquid composition, whereas inother embodiments, the composition is a solid or semi-solid composition.For example, the composition may be a liquid composition suitable forintravenous administration or administration by infusion, or thecomposition may be a solid or semi-solid composition, such as a pill,tablet, or capsule, suitable for oral administration.

Furthermore, the present invention includes methods and compositionsrelated to the use of a halogen compound in combination with one or moreadditional active agents for the treatment or prevention of any of thediseases, conditions or injuries described herein, including thoseassociated with or resulting from ischemia, hypoxia, or reperfusion.These methods include providing to a subject a composition comprising ahalogen compound in combination with an additional compositioncomprising the one or more additional active agent, as well as methodsthat include providing to the subject a single composition comprisingboth the halogen compound and the one or more additional active agent.The present invention contemplates the use of a wide variety ofadditional active agents, including, e.g., chalcogenides, such assulfides and selenides, as well as other agents. In particularembodiments, the composition comprises a halogen compound and anotheractive agent used to treat or prevent chronic heart failure. Thecompositions may be formulated for a variety of different routes ofadministration, including but not limited to, intravenousadministration, administration by infusion, or oral administration.

In certain embodiments, the present invention includes compositions thatcomprise a carrier, wherein a halogen compound is associated with thecarrier. Additional active agents, such as chalcogenides may also beassociated with the carrier. In one embodiment, the carrier is albuminor a related polypeptide, plasma, serum, alpha-2-macroglobulin, orimmunoglobulin.

The present invention is also based, in part, on the surprising andunexpected finding that glutathione stabilizes or prevents oxidation ofchalcogenides, including selenide. Accordingly, the present inventionincludes compositions (e.g., pharmaceutical compositions) comprisingglutathione and a chalcogenide (such as selenide), optionally incombination with a halogen compound (such as iodide), which may be usedto treat or prevent diseases or injuries, including, e.g., thoseassociated with hypoxia, ischemia or reperfusion. In particularembodiments, the composition is formulated for intravenousadministration, administration by infusion, or oral administration. Thecompositions of the present invention are particularly advantageous,because the glutathione inhibits oxidation of the chalcogenide, thusmaking the composition more stable are extending its shelf life.

The present invention also includes unit dosage forms of a compositioncomprising an effective amount of a chalcogenide and glutathione, whichare useful, inter alia, in treating a variety of diseases, conditions,and injuries. The composition may further comprise an effective amountof a halogen compound. In particular embodiments, the unit dosage formscomprise or constitute a pre-measured effective amount of achalcogenide, e.g., selenide or sulfide, which is advantageous fordelivering an appropriate effective amount of a chalcogenide to asubject, particularly during emergency intervention at site of injury orduring transfer of a subject. In certain embodiments, the composition isa liquid composition, whereas in other embodiments, the composition is asolid or semi-solid composition. For example, the composition may be aliquid composition suitable for intravenous administration oradministration by infusion, or the composition may be a solid orsemi-solid composition, such as a pill, tablet, or capsule, suitable fororal administration. The present invention is also based, in part, onthe finding that glutathione stabilizes or prevents oxidation ofhalogens, including iodine compounds, e.g., iodide and iodate.Accordingly, the present invention includes compositions (e.g.,pharmaceutical compositions) comprising glutathione and a halogencompound (such as an iodine compound, e.g., iodide or iodate),optionally in combination with another active agent, e.g., achalcogenide compound (such as sulfide or selenide), which may be usedto treat or prevent diseases or injuries, including, e.g., thoseassociated with hypoxia, ischemia or reperfusion. In particularembodiments, the another active agent is one used to treat or preventchronic heart failure. In certain embodiments, the composition comprisesa halogen compound, glutathione, and another active agent used to treator prevent heart failure. In particular embodiments, the composition isformulated for intravenous administration, administration by infusion,or oral administration. The compositions of the present invention areparticularly advantageous, because the glutathione inhibits oxidation ofthe halogen compounds (and also the chalcogenide, if present), thusmaking the composition more stable are extending its shelf life.

The present invention also includes unit dosage forms of a compositioncomprising an effective amount of a halogen compound and glutathione,which are useful, inter alia, in treating a variety of diseases,conditions, and injuries. The composition may further comprise aneffective amount of a chalcogenide and/or another active agent used totreat chronic heart failure. In particular embodiments, the unit dosageforms comprise or constitute a pre-measured effective amount of ahalogen compound, e.g., an iodine compound, such as iodide or iodate,which is advantageous for delivering an appropriate effective amount ofa halogen compound to a subject, particularly during emergencyintervention at site of injury or during transfer of a subject. Incertain embodiments, the composition is a liquid composition, whereas inother embodiments, the composition is a solid or semi-solid composition.For example, the composition may be a liquid composition suitable forintravenous administration or administration by infusion, or thecomposition may be a solid or semi-solid composition, such as a pill,tablet, or capsule, suitable for oral administration.

The present invention also includes unit dosage forms of a compositioncomprising an effective amount of a halogen compound and another activeagent used to treat or prevent chronic heart failure, which are useful,inter alia, in treating or preventing chronic heart failure, e.g.,following heart attack. The composition may further comprise aneffective amount of a chalcogenide and/or glutathione. In particularembodiments, the unit dosage forms comprise or constitute a pre-measuredeffective amount of a halogen compound, e.g., iodide, and other activeagent that is advantageous for delivering an appropriate effectiveamount of the halogen compound and other active agent to a subject,e.g., a subject suffering from chronic heart failure, or following heartattack. In certain embodiments, the composition is a liquid composition,whereas in other embodiments, the composition is a solid or semi-solidcomposition. For example, the composition may be a liquid compositionsuitable for intravenous administration or administration by infusion,or the composition may be a solid or semi-solid composition, such as apill, tablet, or capsule, suitable for oral administration.

The present invention is also based, in part, on the discovery thatselenide, iodide and other compounds that inhibit thyroid hormoneproduction, activity, or uptake, including, e.g., goitrogens, impede orinhibit peroxide formation at the site of injury or disease, thuspreventing the harmful effects of peroxide and, thus, preventing,inhibiting, or reducing said injury or disease. Accordingly, the presentinvention includes methods for treating or preventing any of theinjuries and diseases described herein in a subject by providing to thesubject a goitrogen, or an active agent that inhibits or impedes theformation of peroxide at the site of injury or disease. In particularembodiments, such active agent is iodide or selenide.

Without wishing to be bound by any particular theory, it is believedthat the inhibition of thyroid hormone production by iodine is due, atleast in part, to the Wolff Chaikoff effect. The Wolff-Chaikoff effectis a reduction in thyroid hormone levels caused by ingestion of a largeamount of iodine. It is considered to be an autoregulatory phenomenonthat inhibits organification (oxidation of iodide) in the thyroid gland,the formation of thyroid hormones inside the thyroid follicle, and therelease of thyroid hormones into the bloodstream. This becomes evidentsecondary to elevated levels of circulating iodide. The Wolff-Chaikoffeffect lasts several days (around 10 days), after which it is followedby resumption of normal organification of iodine and normal thyroidperoxidase function, which is believed to occur because of decreasedinorganic iodine concentration secondary to down-regulation ofsodium-iodide symporter (NIS) on the basolateral membrane of the thyroidfollicular cell. By impeding thyroid hormone production, activity, orlevels at the site of injury or disease, it is thought that metabolicactivity is reduced, or hypermetabolic activity is inhibited, resultingin reduced damage to the injured or diseased tissue. While it is unknownhow high iodide levels impede thyroid peroxidase to produce H₂O₂,without wishing to be bound to any particular theory, it is hypothesizedthat iodide donates electrons to hydrogen peroxide to make water, whichhas been incorrectly interpreted as inhibition of thyroperoxidase.

The present invention is also directed to the use of iodide or otherelemental reducing agents (ERAs), such as sulfide or selenide, to treator prevent any of the diseases, disorder, conditions, or injuriesdescribed herein. In addition, the present invention is also directed,in part, to methods of using halogen compounds and/or chalcogenides toinhibit, prevent or reduce an immune response in a subject. As describedherein, halogen compounds, e.g., iodide, and chalcogenides, e.g.,selenide, can inhibit or reduce reactive oxygen species. Accordingly,they may be used to reduce metabolic activity and stress, as well asinflammation and undesirable immune responses, including, e.g., thoseresulting from reactive oxygen species.

Compositions of the present invention, including stable compositionscomprising a reduced form of halogen compound and/or a reduced form of achalcogenide, may be used in any of the various methods of the presentinvention.

Definitions and Abbreviations

Unless otherwise defined herein, scientific and technical terms used inthis application shall have the meanings that are commonly understood bythose of ordinary skill in the art. Generally, nomenclature used inconnection with, and techniques of, chemistry, molecular biology, celland cancer biology, immunology, microbiology, pharmacology, and proteinand nucleic acid chemistry, described herein, are those well-known andcommonly used in the art.

As used herein, the following terms have the meanings ascribed to themunless specified otherwise.

The term “including” is used to mean “including but not limited to.”“Including” and “including but not limited to” are used interchangeably.

The words “a” and “an” denote one or more, unless specifically noted.

By “about” is meant a quantity, level, value, number, frequency,percentage, dimension, size, amount, weight or length that varies by asmuch as 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1% to a referencequantity, level, value, number, frequency, percentage, dimension, size,amount, weight or length. In any embodiment discussed in the context ofa numerical value used in conjunction with the term “about,” it isspecifically contemplated that the term about can be omitted.

Unless the context requires otherwise, throughout the presentspecification and claims, the word “comprise” and variations thereof,such as, “comprises” and “comprising” are to be construed in an open,inclusive sense, that is as “including, but not limited to”.

By “consisting of” is meant including, and limited to, whatever followsthe phrase “consisting of.” Thus, the phrase “consisting of” indicatesthat the listed elements are required or mandatory, and that no otherelements may be present.

By “consisting essentially of” is meant including any elements listedafter the phrase, and limited to other elements that do not interferewith or contribute to the activity or action specified in the disclosurefor the listed elements. Thus, the phrase “consisting essentially of”indicates that the listed elements are required or mandatory, but thatother elements are optional and may or may not be present depending uponwhether or not they affect the activity or action of the listedelements.

Reference throughout this specification to “one embodiment” or “anembodiment” means that a particular feature, structure or characteristicdescribed in connection with the embodiment is included in at least oneembodiment of the present invention. Thus, the appearances of thephrases “in one embodiment” or “in an embodiment” in various placesthroughout this specification are not necessarily all referring to thesame embodiment. Furthermore, the particular features, structures, orcharacteristics may be combined in any suitable manner in one or moreembodiments.

An “increased” or “enhanced” amount is typically a “statisticallysignificant” amount, and may include an increase that is 1.1, 1.2, 1.3,1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10,15, 20, 30, 40, or 50 or more times (e.g., 100, 500, 1000 times)(including all integers and decimal points in between and above 1, e.g.,2.1, 2.2, 2.3, 2.4, etc.) an amount or level described herein.

A “decreased” or “reduced” or “lesser” amount is typically a“statistically significant” amount, and may include a decrease that isabout 1.1, 1.2, 1.3, 1.4, 1.5, 1.6 1.7, 1.8, 1.9, 2, 2.5, 3, 3.5, 4,4.5, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, or 50 or more times (e.g., 100,500, 1000 times) (including all integers and decimal points in betweenand above 1, e.g., 1.5, 1.6, 1.7, 1.8, etc.) an amount or leveldescribed herein.

A “composition” can comprise an active agent, e.g., a halogen compoundand/or a chalcogenide, and a carrier, inert or active, e.g., apharmaceutically acceptable carrier, diluent or excipient. A compositionmay be a pharmaceutical composition. In particular embodiments, thecompositions are sterile, substantially free of endotoxins or non-toxicto recipients at the dosage or concentration employed.

“Pharmaceutically acceptable carrier, diluent or excipient” includeswithout limitation any adjuvant, carrier, excipient, glidant, sweeteningagent, diluent, preservative, dye/colorant, flavor enhancer, surfactant,wetting agent, dispersing agent, suspending agent, stabilizer, isotonicagent, solvent or emulsifier which has been approved by the UnitedStates Food and Drug Administration as being acceptable for use inhumans or domestic animals.

The term “biological matter” refers to any living biological material,including cells, tissues, organs, and/or organisms, and any combinationthereof. It is contemplated that the methods of the present inventionmay be practiced on a part of an organism (such as in cells, in tissue,and/or in one or more organs), whether that part remains within theorganism or is removed from the organism, or on the whole organism.Moreover, it is contemplated in the context of cells and tissues thathomogenous and heterogeneous cell populations may be the subject ofembodiments of the invention. The term “in vivo biological matter”refers to biological matter that is in vivo, i.e., still within orattached to an organism. Moreover, the term “biological matter” will beunderstood as synonymous with the term “biological material.” In certainembodiments, it is contemplated that one or more cells, tissues, ororgans is separate from an organism. The term “isolated” can be used todescribe such biological matter. It is contemplated that the methods ofthe present invention may be practiced on in vivo and/or isolatedbiological matter.

The terms “mammal” and “subject” includes human and non-human mammals,such as, e.g., a human, mouse, rat, rabbit, monkey, cow, hog, sheep,horse, dog, and cat.

“Pharmaceutically acceptable salts” include sulfate, citrate, acetate,oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acidphosphate, lsomcotinate, lactate, salicylate, acid citrate, tartrate,oleate, tannate, pantothenate, bitartrate, ascorbate, succinate,maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate,formate, benzoate, glutamate, methanesulfonate, ethanesulfonate,benzenesulfonate, p-toluenesulfonate, camphorsulfonate, pamoate,phenylacetate, trifluoroacetate, acrylate, chlorobenzoate,dimtrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate,o-acetoxybenzoate, naphthalene-2-benzoate, isobutyrate, phenylbutyrate,alpha-hydroxybutyrate, butyne-1,4-dicarboxylate,hexyne-1,4-dicarboxylate, caprate, caprylate, cinnamate, glycollate,heptanoate, hippurate, malate, hydroxymaleate, malonate, mandelate,mesylate, mcotinate, phthalate, teraphthalate, propiolate, propionate,phenylpropionate, sebacate, suberate, p-bromobenzenesulfonate,chlorobenzenesulfonate, ethylsulfonate, 2-hydroxyethylsulfonate,methylsulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate,naphthalene-1,5-sulfonate, xylenesulfonate, and tartarate salts. Theterm “pharmaceutically acceptable salt” also refers to a salt of anantagonist of the present invention having an acidic functional group,such as a carboxylic acid functional group, and a base. Suitable basesinclude, but are not limited to, hydroxides of alkali metals such assodium, potassium, and lithium, hydroxides of alkaline earth metal suchas calcium and magnesium, hydroxides of other metals, such as aluminumand zinc, ammonia, and organic amines, such as unsubstituted orhydroxy-substituted mono-, di-, or tri-alkylamines, dicyclohexylamine,tributylamine, pyridine, N-methyl, N-ethylamine, diethylamine,triethylamine, mono-, bis-, or tris-(2-OH-lower alkylamines), such asmono-, bis-, or tris-(2-hydroxyethyl)amine, 2-hydroxy-tert-butylamine,or tris-(hydroxymethyl)methylamine, N,N-di-lower alkyl-N-(hydroxyl-loweralkyl)-amines, such as N,N-dimethyl-N-(2-hydroxyethyl)amine ortri-(2-hydroxyethyl)amine, N-methyl-D-glucamine, and amino acids such asarginine, lysine, and the like. The term “pharmaceutically acceptablesalt” also includes a hydrate of a compound of the invention.

The terms “tissue” and “organ” are used according to their ordinary andplain meanings Though tissue is composed of cells, it will be understoodthat the term “tissue” refers to an aggregate of similar cells forming adefinite kind of structural material. Moreover, an organ is a particulartype of tissue. In certain embodiments, the tissue or organ is“isolated,” meaning that it is not located within an organism.

The terms “hypoxia” and “hypoxic” refer to an environment with levels ofoxygen below normal. Hypoxia occurs when the normal physiologic levelsof oxygen are not supplied to a cell, tissue, or organ. “Normoxia”refers to normal physiologic levels of oxygen for the particular celltype, cell state or tissue in question. “Anoxia” is the absence ofoxygen. “Hypoxic conditions” are those leading to cellular, organ ororganismal hypoxia. These conditions depend on cell type, and on thespecific architecture or position of a cell within a tissue or organ, aswell as the metabolic status of the cell. For purposes of the presentinvention, hypoxic conditions include conditions in which oxygenconcentration is at or less than normal atmospheric conditions, that isless that 20.8, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6,5, 4, 3, 2, 1, 0.5, 0%. Alternatively, these numbers could represent thepercent of atmosphere at 1 atmosphere of pressure (101.3 kPa). “Anoxia”is the absence of oxygen. An oxygen concentration of zero percentdefines anoxic conditions. Thus, hypoxic conditions include anoxicconditions, although in some embodiments, hypoxic conditions of not lessthan 0.5% are implemented. As used herein, “normoxic conditions”constitute oxygen concentrations of around 20.8% or higher.

The term “buffer” as used herein denotes a pharmaceutically acceptableexcipient, which stabilizes the pH of a pharmaceutical preparation.Suitable buffers are well known in the art. Suitable pharmaceuticallyacceptable buffers include but are not limited to acetate-buffers,histidine-buffers, citrate-buffers, succinate-buffers, tris-buffers andphosphate-buffers. In certain embodiments, the concentration of thebuffer is from about 0.01 mM to about 1000 mM, about 0.1 mM to bout 1000mM, about 0.1 mM to about 500 mM, about 0.1 to about 200 mM, about 0.1to about 100 mM, about 1 mM to about 1000 mM, about 1 mM to about 500mM, about 1 mM to about 200 mM, about 1 mM to about 100 mM, about 1 mMto about 50 mM, about 2 mM to about 60 mM, about 4 mM to about 60 mM, orabout 4 mM to about 40 mM, about 5 mM to about 20 mM, or about 5 mM toabout 25 mM.

Pharmaceutically acceptable “cryoprotectants” are known in the art andinclude without limitation, e.g., sucrose, trehalose, and glycerol.Pharmaceutically acceptable cryoprotectants provide stability protectionof compositions, or one or more active ingredients therein, from theeffects of freezing and/or lyophilization.

The term “tonicity agent” or “tonicity modifier” as used herein denotespharmaceutically acceptable agents used to modulate the tonicity of acomposition. Suitable tonicity agents include, but are not limited to,sodium chloride, sorbitol, trehalose, potassium chloride, glycerin andany component from the group of amino acids, sugars, as defined hereinas well as combinations thereof. In certain embodiments, tonicity agentsmay be used in an amount of about 1 mM to about 1000 mM, about 1 mM toabout 500 mM, about 5 mM to about 500 mM, about 10 mM to about 450 mM,about 20 mM to about 400 mM, about 50 mM to about 300 mM, about 100 mMto about 200 mM, or about 125 mM to about 175 mM. In certainembodiments, a tonicity agent comprises an amino acid present in acomposition at about 5 mM to about 500 mM.

The term “stabilizer” indicates a pharmaceutical acceptable excipient,which protects the active pharmaceutical ingredient(s) or agents(s)and/or the composition from chemical and/or physical degradation duringmanufacturing, storage and application. Stabilizers include, but are notlimited to, sugars, amino acids, polyols, surfactants, antioxidants,preservatives, cyclodextrines, e.g. hydroxypropyl-β-cyclodextrine,sulfobutylethyl-β-cyclodextrin, β-cyclodextrin, polyethyleneglycols,e.g. PEG 3000, PEG 3350, PEG 4000, PEG 6000, albumin, e.g. human serumalbumin (HSA), bovine serum albumin (BSA), salts, e.g. sodium chloride,magnesium chloride, calcium chloride, and chelators, e.g. EDTA.Stabilizers may be present in the composition in an amount of about 0.1mM to about 1000 mM, about 1 mM to about 500 mM, about 10 to about 300mM, or about 100 mM to about 300 mM.

As used herein, the term “surfactant” refers to a pharmaceuticallyacceptable organic substance having amphipathic structures; namely, itis composed of groups of opposing solubility tendencies, typically anoil-soluble hydrocarbon chain and a water-soluble ionic group.Surfactants can be classified, depending on the charge of thesurface-active moiety, into anionic, cationic, and nonionic surfactants.Surfactants may be used as wetting, emulsifying, solubilizing, anddispersing agents for pharmaceutical compositions and preparations ofbiological materials. In some embodiments of the compositions describedherein, the amount of surfactant is described as a percentage expressedin weight/volume percent (w/v %). Suitable pharmaceutically acceptablesurfactants include, but are not limited to, the group ofpolyoxyethylensorbitan fatty acid esters (Tween), polyoxyethylene alkylethers (Brij), alkylphenylpolyoxyethylene ethers (Triton-X),polyoxyethylene-polyoxypropylene copolymer (Poloxamer, Pluronic), orsodium dodecyl sulphate (SDS). Polyoxyethylenesorbitan-fatty acid estersinclude polysorbate 20, (sold under the trademark Tween 20™) andpolysorbate 80 (sold under the trademark Tween 80™).Polyethylene-polypropylene copolymers include those sold under the namesPluronic® F68 or Poloxamer 188™. Polyoxyethylene alkyl ethers includethose sold under the trademark Brij™ Alkylphenolpolyoxyethylene ethersinclude those sold under the tradename Triton-X. Polysorbate 20 (Tween20™) and polysorbate 80 (Tween 80™) are generally used in aconcentration range of about 0.001% w/v to about 1% w/v or about 0.002%w/v to about 0.1% w/v of the total volume of the composition, oralternatively of about 0.003% w/v to about 0.007% w/v. In someembodiments, Tween 80™ is used at about 0.003% w/v, about 0.004% w/v,about 0.0045% w/v, about 0.005% w/v, about 0.0055% w/v, about 0.006% w/vor about 0.007% w/v. In some embodiments, Tween 80™ is used at about0.005% w/v. In this aspect, “w/v” intends the weight of surfactant pertotal volume of the composition.

A “lyoprotectant” refers to a pharmaceutically acceptable substance thatstabilizes a protein, nucleic acid or other active pharmaceuticalingredient(s) or agent(s) during lyophilization. Examples oflyoprotectants include, without limitation, sucrose, trehalose ormannitol.

A “polyol” refers to an alcohol containing multiple hydroxyl groups, ora sugar alcohol. A sugar alcohol is a hydrogenated form of carbohydrate,whose carbonyl group (aldehyde or ketone, reducing sugar) has beenreduced to a primary or secondary hydroxyl group (hence the alcohol).Sugar alcohols have the general formula H(HCHO)_(n+1)H, whereas sugarshave H(HCHO)_(n)HCO.

An “antioxidant” refers to a molecule capable of slowing or preventingthe oxidation of other molecules. Antioxidants are often reducingagents, chelating agents and oxygen scavengers such as thiols, ascorbicacid or polyphenols. Non-limiting examples of antioxidants includeascorbic acid (AA, E300), thiosulfate, methionine, tocopherols (E306),propyl gallate (PG, E310), tertiary butylhydroquinone (TBHQ), butylatedhydroxyanisole (BHA, E320) and butylated hydroxytoluene (BHT, E321).

A “preservative” is a natural or synthetic chemical that is added toproducts such as foods, pharmaceutical compositions, paints, biologicalsamples, wood, etc. to prevent decomposition by microbial growth or byundesirable chemical changes. Preservative additives can be used aloneor in conjunction with other methods of preservation. Preservatives maybe antimicrobial preservatives, which inhibit the growth of bacteria andfungi, or antioxidants such as oxygen absorbers, which inhibit theoxidation of constituents. Examples of antimicrobial preservativesinclude benzalkonium chloride, benzoic acid, chlorohexidine, glycerin,phenol, potassium sorbate, thimerosal, sulfites (sulfur dioxide, sodiumbisulfite, potassium hydrogen sulfite, etc.) and disodium EDTA. Otherpreservatives include those commonly used in parenteral proteincompositions such as benzyl alcohol, phenol, m-cresol, chlorobutanol ormethylparaben.

As used in the specification and appended claims, unless specified tothe contrary, the following terms have the meaning indicated:

“Mammal” includes humans and both domestic animals such as laboratoryanimals and household pets, (e.g., cats, dogs, swine, cattle, sheep,goats, horses, and rabbits), and non-domestic animals such as wildlifeand the like.

“Optional” or “optionally” means that the subsequently described eventof circumstances may or may not occur, and that the description includesinstances where said event or circumstance occurs and instances in whichit does not.

“Pharmaceutical composition” refers to a formulation of a compound and amedium generally accepted in the art for the delivery of thebiologically active compound to mammals, e.g., humans. Such a medium mayinclude any pharmaceutically acceptable carriers, diluents or excipientstherefore.

“Sulfide” refers to sulfur in its −2 valence state, e.g., either as H₂Sor as a salt thereof (e.g., NaHS, Na2S, etc.).

“Selenide” refers to selenium in its −2 valence state, e.g., either asH₂Se or as a salt thereof (e.g., NaHSe, Na₂Se, etc.).

“Iodide” and “a reduced form of iodide” both refer to iodide, which hasa −1 valence state (e.g., NaI). “A reduced form of iodine” includesiodide.

“Chalcogenide” or “chalcogenide compounds” refers to compoundscontaining a chalcogen element, i.e., those in Group 6 of the periodictable, but excluding oxides. These elements are sulfur (S), selenium(Se), tellurium (Te) and polonium (Po). Specific chalcogenides and saltsthereof include, but are not limited to: H₂S, Na₂S, NaHS, K₂S, KHS,Rb₂S, CS₂S, (NH4)₂S, (NH₄)HS, BeS, MgS, CaS, SrS, BaS, H₂Se, Na₂Se,NaHSe, K₂Se, KHSe, Rb₂Se, CS2Se, (NH4)₂Se, (NH₄)HSe, BeSe, MgSe, CaSe,SrSe, PoSe and BaSe.

As used herein, “oxidation product” refers to products that result fromoxidation of a compound, e.g., a chalcogenide or halide compound,including, e.g., sulfite, sulfate, thiosulfate, polysulfides,dithionate, polythionate, elemental sulfur, selenite, selenate,thioselenate, polyselenides, and elemental selenium. Such oxidationproducts could occur as a result of processing, manufacturing or storage(e.g., by oxidation).

A used herein, “goitrogen” refers to a substance that inhibits theproduction of thyroid hormone by the body. While the mechanism is notentirely understood, it is believed that certain goitrogens suppress thefunction of the thyroid gland by interfering with iodine uptake ororganification (synthesis of thyroid hormone), which can, as a result,cause an enlargement of the thyroid, i.e., a goiter, whereas othergoitrogens may exert their effect through other mechanisms. Chemicalsthat have been shown to have goitrogenic effects include:

sulfadimethoxine, propylthiouracil, potassium perchlorate, and iopanoicacid;

some oxazolidines such as goitrin;

thiocyanate, e.g., thiocyanate overload in Central Africa, especially ifalso in conjunction with selenium deficiency. Reliance on cassava as acarbohydrate provides a source of thiocyanate in some areas;

ions such as thiocyanate and perchlorate, which decrease iodide uptakeby competitive inhibition and, as a consequence of reduced thyroxine andtriiodothyronine secretion by the gland, cause, at low doses, anincreased release of thyrotropin (by reduced negative feedback), whichthen stimulates the gland;

amiodarone, which inhibits peripheral conversion of thyroxine totriiodothyronine; also interferes with thyroid hormone action;

lithium inhibits thyroid hormone release;

methimazole; and

phenobarbitone, phenytoin, carbamazepine, rifampin induce metabolicdegradation of triiodothyronine (T₃) and thyroxine (T₄).

“Therapeutically effective amount” refers to that amount of a compoundor composition of the invention that, when administered to a biologicalmaterial, e.g., a mammal, preferably a human, is sufficient to effecttreatment, as defined below, of a disease, injury, or condition in thebiological material, e.g., mammal, preferably a human. The amount of acompound or composition of the invention which constitutes a“therapeutically effective amount” will vary depending on the compoundor composition, the disease, injury or condition and its severity, themanner of administration, and the age of the biological material, e.g.,mammal, to be treated, but can be determined routinely by one ofordinary skill in the art having regard to his own knowledge and to thisdisclosure.

“Treating” or “treatment” as used herein covers the treatment of thedisease, injury, or condition of interest, e.g., tissue injury, in abiological material, e.g., mammal, preferably a human, having thedisease or condition of interest, and includes: (i) preventing orinhibiting the disease, injury, or condition from occurring in abiological material, e.g., mammal, in particular, when such mammal ispredisposed to the condition but has not yet been diagnosed as havingit; (ii) inhibiting the disease, injury, or condition, i.e., arrestingits development; (iii) relieving the disease, injury, or condition,i.e., causing regression of the disease or condition; or (iv) relievingthe symptoms resulting from the disease, injury, or condition. As usedherein, the terms “disease,” “disorder,” and “condition” may be usedinterchangeably. As used herein, the term “injury” includesunintentional injuries and intentional injuries, including injuries thatoccur, “at the hand of man,” including injuries associated with medicalprocedures, such as surgeries and transplantations.

“Chronic heart failure” (CHF), also referred to as congestive heartfailure (CHF) or congestive cardiac failure (CCF), occurs when the heartis unable to pump sufficiently to maintain blood flow to meet the needsof the body. Common causes of CHF include coronary artery diseaseincluding a previous myocardial infarction (heart attack), high bloodpressure, atrial fibrillation, valvular heart disease, andcardiomyopathy. These cause heart failure by changing either thestructure or the functioning of the heart. The two main types of heartfailure are: (1) heart failure due to left ventricular dysfunction,wherein the ability of the left ventricle to contract is affected, whichis also referred to as systolic heart failure; and (2) heart failurewith normal ejection fraction, wherein the heart's ability to relax isaffected, which is also referred to as diastolic heart failure.

Halogen Compounds

The present invention relates to halogen compounds, said compoundscomprising any element included in Group 17 of the periodic table. Anyform of halogen compound may be used according to the present invention,including, e.g., hydrogen halides, metal halides, interhalogencompounds, organohalogen compounds, and polyhalogenated compounds. Insome embodiments, a halogen compound refers to any compound containingFluorine, Chlorine, Bromine, Iodine, Astatine, or Ununseptium. Inparticular embodiments, the present invention relates to halogencompounds in a reduced form, e.g., iodide.

Fluorine (F), the lightest halogen, is the non-metal element with atomicnumber 9. Under standard pressure and temperature it exists as adiatomic gas F₂. Fluorine is the most chemically reactive element,reacting with all other elements except oxygen, helium, neon, andkrypton. It is also the most electronegative element, thus attractingelectrons more strongly than all other elements. There are 11 fluorineisotopes with known half-lives, said isotopes having mass numbersranging from 15 to 25. Natural Fluorine, however, consists of one stableisotope, ¹⁹F.

Chlorine (Cl), the second lightest halogen, is the non-metal elementwith atomic number 17. Under standard pressure and temperature it existsas a diatomic gas F₂. Chlorine is the element with the highest electronaffinity, and the third highest electronegativity. There are 16 chlorineisotopes with known half-lives, said isotopes having mass numbersranging from 31 to 46. Naturally occurring chlorine is a mixture of twostable isotope ³⁵Cl and ³⁷Cl, existing in natural abundance ratios ofapproximately 3:1.

Bromine (Br), the third lightest halogen, is the non-metal element withatomic number 35. Under standard pressure and temperature it exists as adiatomic liquid Br₂. There are 26 bromine isotopes with know half-lives,said isotopes having mass numbers ranging from 68 to 94. Naturallyoccurring bromine is a mixture of two stable isotope ⁷⁹Cl and ⁸¹Cl,existing in natural abundance ratios of approximately 1:1.

Iodine (I), the second heaviest natural halogen, is the non-metalelement with atomic number 53. Under standard pressure and temperatureit exists as a solid diatomic I₂ molecule. There are 34 iodine isotopeswith known half-lives, said isotopes having mass numbers ranging from108 to 144. Natural iodine, however, consists of one stable isotope,¹²⁷I.

Astatine, the heaviest natural halogen, is a highly radioactive thenon-metal element with atomic number 85. It decays so rapidly (longesthalf-life less than 12 hours) that its properties are not known withgreat certainty. It is debated if astatine exists as a diatomic At₂molecule, as this form has never actually been observed. Astatine canreact with hydrogen to form hydrogen astatide, and it is predicted toreact with metals such as sodium to form salts. There are 37 knownastatine isotopes, all of which are radioactive, with very shorthalf-lives. Said isotopes have mass numbers ranging from 207 to 221. Nostable isotopes of astatine exist.

In various embodiments, compositions and methods of the presentinvention comprise one or more halogen compounds, such as various formsof iodine or bromine.

In one embodiment, the present invention relates to a halogen compoundcontaining iodine. In particular embodiments, the halogen compound is areduced form of iodine, such as iodide. Certain embodiments may comprisean iodine-containing halogen compound that is an iodide, iodate,organoiodide, periodate, or periodinane.

In some embodiments, said halogen compound is an iodide comprising oneor more compounds from the non-limiting list of Aluminium iodide,Aluminium monoiodide, Ammonium iodide, Antimony triiodide, Arsenicdiiodide, Arsenic triiodide, Barium iodide, Beryllium iodide,Bismuth(III) iodide, Boron triiodide, Cadmium iodide, Caesium iodide,Calcium iodide, Candocuronium iodide, Carbon tetraiodide, Cobalt(II)iodide, Coccinite, Copper(I) iodide, DiOC6, Diphosphorus tetraiodide,Dithiazanine iodide, Echothiophate, Einsteinium(III) iodide,Eschenmoser's salt, Ethylenediamine dihydroiodide, Gallium(III) iodide,GelGreen, GelRed, Germanium iodide, Gold monoiodide, Gold triiodide,Hydrogen iodide, Iodine oxide, Iodomethylzinc iodide, Iodosilane,Iron(II) iodide, Lead(II) iodide, Lithium iodide, Magnesium iodide,Manganese(II) iodide, Mercury(I) iodide, Mercury(II) iodide, Nickel(II)iodide, Nitrogen triiodide, Palladium(II) iodide, Phosphorus triiodide,Polyiodide, Potassium iodide, Potassium tetraiodomercurate(II),Propidium iodide, Rubidium iodide, Rubidium silver iodide, Samarium(II)iodide, Silicon tetraiodide, Silver iodide, Sodium iodide, Strontiumiodide, Tellurium iodide, Tellurium tetraiodide, Terbium(III) iodide,Tetraethylammonium iodide, Thallium triiodide, Thallium(I) iodide,Thorium(IV) iodide, Tibezonium iodide, Tiemonium iodide, Tin(II) iodide,Tin(IV) iodide, Titanium tetraiodide, Triiodide, Trimethylsilyl iodide,Trimethylsulfoxonium iodide, Uranium pentaiodide, Uranium tetraiodide,Uranium triiodide, Vanadium(III) iodide, Zinc iodide, and Zirconium(IV)iodide.

In particular embodiments, said halogen compound is an iodide comprisingsodium iodide, potassium iodide, hydrogen iodide, calcium iodide, orsilver iodide.

In some embodiments, said halogen compound is an iodate comprising oneor more compounds from the non-limiting list of Calcium iodate, Iodicacid, Potassium iodate, Seeligerite, Silver iodate, and Sodium iodate.

In particular embodiments, said halogen compound is an iodate comprisingsodium iodate, potassium iodate, calcium iodate, or silver iodate.

In some embodiments, said halogen compound is an organoiodide comprisingone or more compounds from the non-limiting list of ²⁵I-NBF, ²⁵I-NBMD,²⁵I-NBOH, ²⁵I-NBOMe, 2C-I, 5, 5-I-R91150, Acetrizoic acid, Adipiodone,Adosterol, Altropane, AM-1241, AM-2233, AM-630, AM-679 (cannabinoid),AM-694, AM251, Amiodarone, Benziodarone, Bromoiodomethane, Budiodarone,Butyl iodide, Carbon tetraiodide, Chiniofon, Chloroiodomethane,Clioquinol, Diatrizoic acid, Diiodohydroxypropane,Diiodohydroxyquinoline, Diiodomethane, 2,5-Dimethoxy-4-iodoamphetamine,Domiodol, Erythrosine, Ethyl iodide, Ethyl iodoacetate, Fialuridine,Fluoroiodomethane, Haloprogin, Herapathite, IAEDANS, Ibacitabine, IDNNA,Idoxifene, Idoxuridine, Iniparib, Iobenguane, Iobenzamic acid,Iobitridol, Iocarmic acid, Iocetamic acid, Iodamide, Iodixanol,Iodoacetamide, Iodoacetic acid, Para-Iodoamphetamine, Iodobenzamide,Iodobenzene, 2-Iodobenzoic acid, 19-Iodocholesterol, Iodocyanopindolol,Iodoform, 1-Iodomorphine, Iodophenol, Iodophenpropit, 4-Iodopropofol,Iodopropynyl butylcarbamate, Iodotrifluoroethylene, Iodoxamic acid,2-Iodoxybenzoic acid, Iofetamine (1231), Ioflupane (1231), Ioglicicacid, Ioglycamic acid, Iomazenil, Iomeprol, Iopamidol, Iopanoic acid,Iopentol, Iopromide, Iopydol, Iotrolan, Iotroxic acid, Ioversol,Ioxaglic acid, Ioxilan, Ipodate sodium, Isopropyl iodide, Methiodal,Methyl iodide, Metrizamide, Metrizoic acid, Pentafluoroethyl iodide,Plakohypaphorine, N-Propyl iodide, Propyliodone, Rafoxanide, Rosebengal, RTI-121, RTI-229, RTI-353, RTI-55, SB-258,585, Sodiumacetrizoate, Tiratricol, Trifluoroiodomethane, and Tyropanoic acid.

In particular embodiments, said halogen compound is an organoiodide.Organoiodine compounds are organic compounds that contain one or morecarbon-iodine bonds. Almost all organoiodine compounds feature iodideconnected to one carbon center. These are usually classified asderivatives of I⁻. Some organoiodine compounds feature iodine in higheroxidation states. Organoiodine compounds, often used as disinfectants orpesticides, include, e.g., iodoform (CHI₃), methylene iodide (CH₂I₂),and methyl iodide (CH₃I). In particular embodiment, the organoiodide isa polyiodoorganic compound. Polyiodoorganic compounds are sometimesemployed as X-ray contrast agents, in fluoroscopy, a type of medicalimaging. A variety of such polyiodoorganic compounds are availablecommercially; many are derivatives of 1,3,5-triiodobenzene and containabout 50% by weight iodine. In certain embodiments, the agent is solublein water, non-toxic and/or readily excreted. A representative reagent isIoversol, which has water-solubilizing diol substituents. Otherorganoiodine compounds include but are not limited to the two thyroidhormones thyroxine (“T₄”) and triiodothyronine (“T₃”). Marine naturalproducts are rich sources of organoiodine compounds, including therecently discovered plakohypaphorines from the sponge Plakortis simplex.

The present invention also includes the use of compounds, e.g., drugcompounds, into which an iodine is incorporate. For example, an iodinemay be incorporated into existing drugs such as N-acetyl cysteine,standard pain relievers, and non-steroidal anti-inflammatory drugs, suchas, e.g., aspirin, ibuprofen and naproxen. Most NSAIDs act asnonselective inhibitors of the enzyme cyclooxygenase (COX), inhibitingboth the cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2)isoenzymes.

In certain embodiments, said halogen compound is a polyiodide. Thepolyiodides are a class of polyhalogen anions composed of entirelyiodine atoms. The most common and simplest member is the triiodide ion,I₃ ⁻. Other known, larger polyiodides include [I₄]²⁻, [I₅]⁻, [I₇]⁻,[I₈]²⁻, [I₉]⁻, [I₁₀]²⁻, [I₁₀]⁴⁻, [I₁₁]⁻, [I₁₂]²⁻, [I₁₃]⁻³, [I₁₆]⁻²,[I₂₂]⁻⁴, [I₂₆]³⁻, [I₂₆]⁴⁻, [I₂₈]⁴⁻ and [I₂₉]³⁻. One example of apolyiodide is Lugol's iodine, also called Lugol's solution. Lugol'ssolution is commercially available in different potencies of 1%, 2%, or5% Iodine. The 5% solution consists of 5% (wt/v) iodine (I₂) and 10%(wt/v) potassium iodide (KI) mixed in distilled water and has a totaliodine content of 130 mg/mL. Potassium iodide renders the elementaryiodine soluble in water through the formation of the triiodide (I⁻³)ion. Other names for Lugol's solution are I₂KI (iodine-potassiumiodide); Markodine, Strong solution (Systemic); and Aqueous IodineSolution BCP. Examples of polyiodides, including their ions andcounter-cations are shown in Table 1.

TABLE 1 Polyiodides Anion Counter-cation [I₃]⁻ Cs⁺ [I₄]²⁻ [Cu(NH₃)₄]²⁺[I₅]⁻ [EtMe₃N]⁺ [EtMePh₂N]⁺ [I₇]⁻ [Ag(18aneS₆)]⁺ [I₈]²⁻ [Ni(phen)₃]²⁺[I₉]⁻ [Me₂ ^(i)PrPhN]⁺ [Me₄N]⁺ [I₁₀]²⁻ [Cd(12-crown-4)₂]²⁺ [I₁₁]³⁻[(16aneS₄)PdIPd(16aneS₄)]³⁺ [I₁₂]²⁻ [Ag₂(15aneS₅)₂]²⁺ [Cu(Dafone)₃]²⁺[I₁₃]³⁻ [Me₂Ph₂N]⁺ [I₁₆]²⁻ [Me₂Ph₂N]⁺ [^(i)PrMe₂PhN]⁺ [I₂₂]⁴⁻ [MePh₃P]⁺[I₂₆]³⁻ [Me₃S]⁺ [I₂₆]⁴⁻ DMFc⁺ [I₂₉]³⁻ Cp₂Fe [I₂₂]⁴⁻ [MePh₃P]⁺ [I₂₆]³⁻[Me₃S]⁺ [I₂₆]⁴⁻ DMFc⁺ [I₂₉]³⁻ Cp₂Fe [I₂₂]⁴⁻ [MePh₃P]⁺ [I₂₆]³⁻ [Me₃S]⁺[I₂₆]⁴⁻ DMFc⁺

In one embodiment, the halogen compound is a tincture of iodinesolutions, which comprises or consists of elemental iodine, and iodidesalts dissolved in water and alcohol.

In some embodiments, said halogen compound is a periodate comprising oneor more compounds from the non-limiting list of Dess-Martin periodinane,1, 2-Iodoxybenzoic acid, Periodic acid, Potassium periodate, and Sodiumperiodate.

In particular embodiments, said halogen compound is a periodatecomprising sodium periodate, potassium periodate, calcium periodate, orsilver periodate.

In particular embodiments, said halogen compound is a periodinane.Periodinanes are chemical compounds containing hypervalent iodine. Insome embodiments, said halogen compound is a periodinane comprising oneor more compounds from the non-limiting list of(Bis(trifluoroacetoxy)iodo)benzene, Dess-Martin periodinane, Iodobenzenedichloride, Iodosobenzene, and 2-Iodoxybenzoic acid.

In one embodiment, the halogen compound is an oil-infused iodide oriodine oil infusion.

In one embodiment, the present invention relates to a halogen compoundcontaining bromine. Certain embodiments may comprise abromine-containing halogen compound that is a bromide, bromate,organobromide, or a perbromate.

In some embodiments, said halogen compound is a bromide comprising oneor more compounds from the non-limiting list of Aclidinium bromide,Aluminium bromide, Ammonium bromide, ANNINE-6plus, Antimony tribromide,Arsenic tribromide, Barium bromide, Benzododecinium bromide, Berylliumbromide, Bibenzonium bromide, Bismuth tribromide, Boron tribromide,Bromargyrite, Bromo(tetrahydrothiophene)gold(I),Bromopentaamminecobalt(III) bromide, Bromopentacarbonylrhenium(I),Cadmium bromide, Caesium bromide, Caesium cadmium bromide, Calciumbromide, Cerium(III) bromide, Cetrimonium bromide, Chromium(III)bromide, Cimetropium bromide, Clidinium bromide, Cobalt(II) bromide,Copper(I) bromide, Copper(II) bromide, Cyanogen bromide, Demecariumbromide, Ditellurium bromide, DODAB, Domiphen bromide, EEthidiumbromide, Fazadinium bromide, Fentonium, Gallium(III) bromide, Gold(I)bromide, Gold(III) bromide, Hexafluronium bromide, Hydrobromic acid,Hydrogen bromide, Indium(I) bromide, Indium(III) bromide, Iodinemonobromide, Iron(II) bromide, Iron(III) bromide, Lanthanum(III)bromide, Lead(II) bromide, Lithium bromide, Magnesium bromide,Manganese(II) bromide, Mercury(I) bromide, Mercury(II) bromide, Morphinemethylbromide, Nickel(II) bromide, Niobium bromide, Niobium(V) bromide,Nitrogen tribromide, Nitrosyl bromide, Otilonium bromide, Oxitropiumbromide, Oxyphenonium bromide, Palladium(II) bromide, Pancuroniumbromide, Phosphorus heptabromide, Phosphorus pentabromide, Phosphorustribromide, Pifithrin, Pipecuronium bromide, Platinum(II) bromide,Platinum(IV) bromide, Polonium dibromide, Potassium bromide,Propantheline bromide, Radium bromide, Rubidium bromide, Silicontetrabromide, Silver bromide, Sodium bromide, Strontium bromide,TTantalum(V) bromide, Tellurium tetrabromide, Terbium(III) bromide,Tetrabromoauric acid, Tetrabromomethane, Thallium(I) bromide, Timepidiumbromide, Tin(II) bromide, Tin(IV) bromide, Titanium tetrabromide,Tribromosilane, Triphenylcyclopropenium bromide, Tungsten(V) bromide,Tungsten(VI) oxytetrabromide, Uranium pentabromide, Uraniumtetrabromide, Vanadium(III) bromide, Ytterbium(III) bromide,Yttrium(III) bromide, Zinc bromide, and Zirconium(IV) bromide.

In particular embodiments, said halogen compound is a bromide comprisingsodium bromide, potassium bromide, hydrogen bromide, calcium bromide, orsilver bromide.

In some embodiments, said halogen compound is a bromate comprising oneor more compounds from the non-limiting list of Bromic acid, Calciumbromate, Potassium bromate, Silver bromate, Sodium bromate, andStrontium bromate.

In some embodiments, said halogen compound is an organobromidecomprising one or more compounds from the non-limiting list of2-Bromobutyric acid, 25B-NBOMe, 2C-B, 2C-B-BZP, 2C-B-FLY, 2CB-Ind,2CBCB-NBOMe, 2CBFly-NBOMe, 66-Br-APB, Acecarbromal, Ageliferin, Allylbromide, AM-087, Ambroxol, Arbidol, AS-8112, BCDMH, Benzbromarone,Benzyl bromide, Bibrocathol, Brallobarbital, Bretazenil, Bretylium,Bretylium for the treatment of ventricular fibrillation, Brimonidine,Brivudine, Brodifacoum, Brodimoprim, Brofaromine, Bromacil,Bromadiolone, Bromadoline, Bromantane, Bromazepam, Bromazine,Bromethalin, Bromfenac, Bromhexine, Brominated flame retardant,Bromisoval, 2-Bromo-1-chloropropane, 4-Bromo-3,5-dimethoxyamphetamine,2-Bromo-4,5-methylenedioxyamphetamine, Bromo-DragonFLY, Bromoaceticacid, Bromoacetone, Bromoacetylalprenololmenthane, 8-Bromoadenosine3′,5′-cyclic monophosphate, Para-Bromoamphetamine, 4-Bromoaniline,Bromoanisole, Bromobenzene, Bromobimane, 1-Bromobutane, 2-Bromobutane,Bromochlorodifluoromethane, Bromochloromethane,Bromochlorosalicylanilide, Bromocresol green, Bromocresol purple,Bromocriptine, Bromocyclohexane, Bromodeoxyuridine,Bromodichloromethane, Bromodifluoroacetyl chloride,Bromodifluoromethane, Bromodiphenylmethane, B cont.Bromoethane,Bromofluoromethane, Bromoform, 3-Bromofuran, 8-Bromoguanosine3′,5′-cyclic monophosphate, 1-Bromohexane, 2-Bromohexane,Bromoiodomethane, Bromomethane, 4-Bromo-N-methylcathinone, Bromopentane,Bromophenol blue, Bromadol, 2-Bromopropane, Bromopyruvic acid,N-Bromosuccinimide, Bromotrifluoromethane, 5-Bromouracil,5-Bromouridine, Bromoxynil, Bromperidol, Brompheniramine,Bromsulphthalein, Bronidox, Bronopol, Brophebarbital, Bropirimine,Brotizolam, Broxaterol, Broxyquinoline, Butallylonal, Tert-Butylbromide, C-8813, Carbromal, Chlorfenapyr, Ciclotizolam, ConvolutindoleA, DBDMH, DBNPA, Decabromodiphenyl ether, Deltamethrin,Desformylflustrabromine, Dexbrompheniramine, Diarylpyrimidines,1,2-Dibromo-3-chloropropane, 1,4-Dibromobenzene, Dibromochloromethane,Dibromodifluoromethane, 1,1-Dibromoethane, 1,2-Dibromoethane,Dibromofluoromethane, Dibromomethane, 1,2-Dibromopropane,1,3-Dibromopropane, Dibromotetrafluoroethane, Dibromotyrosine,Dibrompropamidine, Difethialone, 2,5-Dimethoxy-4-bromoamphetamine, DS-1(drug), Ebrotidine, Embramine, Eosin, Eosin B, Eosin Y, Ethylbromoacetate, Etravirine, FL3 (flavagline), Flubromazolam, Gidazepam,H-89, Halofuginone, Halomon, Halothane, Haloxazolam,Hexabromocyclododecane, Ibrolipim, Imidazenil, Isobromindione, JWH-249,JWH-424, KF-26777, Lonafarnib, Mebroqualone, Merbromin, Meta-DOB,Metaclazepam, Mitobronitol, Mucobromic acid, Narcobarbital,Nelotanserin, Neltenexine, NGD-4715, Nicergoline, 0-806,Octabromodiphenyl ether, Organobromine compound, P7C3, Pamabrom, PEAQX,Pentabromodiphenyl ether, Phenacyl bromide, Phenazepam,2-Phenylethylbromide, Phloxine, Pinaverium, Pindobind, Pipobroman,PNU-282,987, Polybrominated biphenyl, Polybrominated diphenyl ethers,Propallylonal, Propargyl bromide, N-Propyl bromide, Remoxipride,Romifldine, RTI-51, SB-357,134, Sigmodal, SSR-180,711, Stampidine,Surinabant, Surugatoxin, TCB-2, Tetrabromobisphenol A, Tetrabromoethane,Tetrabromoethylene, Tetrabromomethane, TH-302, Tilbroquinol,Tralomethrin, 2,4,6-Tribromoanisole, and Tribromofluoromethane.

In some embodiments, said halogen compound is a perbromate, saidperbromate comprising sodium perbromate, potassium perbromate, hydrogenperbromate, or silver perbromate.

Particular embodiments of the present invention relate to a reduced formof a halogen compound. Many acceptable means of reduction of halogencompounds are possible and known to one skilled in the art. Examples ofreduced forms of halide compounds include, e.g., iodide and bromide,wherein the halogen has a valency of −1. Non-limiting examples ofreduction methods include chemical reduction with electropositiveelemental metals (such as lithium, sodium, magnesium, iron, zinc, andaluminum, e.g.), hydride transfer reagents (such as NaBH₄ and LiAIH_(4.)e/g), or the use of hydrogen gas with a palladium, platinum, or nickelcatalyst.

A particular embodiment of the present invention relates to theadministration of a halogen compound of the type described herein to amammalian subject, said compounds administered in a composition,concentration or formulation that is not significantly toxic to saidmammals, e.g., a pharmaceutical composition. In particular embodiments,a halogen compound known to be toxic to a mammalian subject is excludedfrom the present invention. Thus, in particular embodiments, potassiumiodide is excluded from the present invention. It is furthercontemplated that some embodiments may comprise the administration ofmore than one of said halogen compounds to said mammal, eithersimultaneously or separately, such that the combination of saidcompounds that are not individually significantly toxic are also notsignificantly toxic when combined.

Other compounds comprising a halogen compound or halogen element mayalso be used according to methods of and/or included in compositions ofthe present invention. In some embodiments, said halogen compound is acommercially available substance. In certain embodiments, saidcommercially available substances may include radiological contrastagents, topical iodine preparations, solutions, or drugs. In certainembodiments, said commercially available substance comprises iodine, andmay be selected from the non-limiting list of Diatrizoate, Ipanoic acid,Ipodate, Iothalamate, Metrizamide, Diatrozide, Diiodohydroxyquinolone,Iodine tincture, Povidone iodine, Iodochlorohydroxyquinolone, Iodoformgauze, Saturated potassium iodide (SSKI), Lugol solution, Iodinatedglycerol, Echothiopate iodide, Hydriodic acid syrup, Calcium iodide,Amiodarone, Expectorants, Vitamins containing iodine,Iodochlorohydroxyquinolone, Diiodohydroxyquinolone, Potassium iodide,Benziodarone, Isopropamide iodide, levothyroxine, and Erythrosine. Incertain embodiments, said commercially available substance comprisesbromine, and may be selected from the non-limiting list of Alphagen(brimonidine), Atrovent (Ipratropium), Celexa (citalopram), Combivent(ipratropium bromide), Enablex (darifenacin), Guaifenex DM(dextromethorphan), Razadyne (galantamine), and Spiriva (tiotropium).

Certain compositions and methods of the present invention comprise ahalogen compound, in combination with glutathione and/or a goitrogen. Asdiscussed herein, glutathione can stabilize halogen compounds in areduced form, and goitrogens can inhibit production of thyroid hormone.

Other Active Agents

The present invention further includes the use of other active agents,alone or in combination with a halogen compound, to treat or prevent anyof the diseases, conditions or injuries described herein. A variety ofagents effective in treating or preventing any of these diseases,conditions or injuries have been described or are known in the art, andany of these agents or other agents may be used. In addition, otheragents may be used in combination with a halogen compound according tothe methods of the present invention.

In certain embodiments, an active agent is a chalcogenide or saltthereof. It has been shown that treatment with a chalcogenide inducesstasis of biological matter and protects biological matter from hypoxicand ischemic injury. In these studies, it was demonstrated that sulfidecan reduce metabolism and protect mice and rats from hypoxic injuries(PCT Publication No. WO2005/041655). Compounds containing a chalcogenelement, i.e., those in Group 6 of the periodic table, but excludingoxides, are commonly termed “chalcogenides” or “chalcogenide compounds.”These elements are sulfur (S), selenium (Se), tellurium (Te) andpolonium (Po). Common chalcogenides contain one or more of S, Se, andTe, in addition to other elements. Specific chalcogenides and saltsthereof include, but are not limited to: H₂S, Na₂S, NaHS, K₂S, KHS,Rb₂S, CS₂S, (NH₄)₂S, (NH₄)HS, BeS, MgS, CaS, SrS, BaS, H₂Se, Na₂Se,NaHSe, K₂Se, KHSe, Rb₂Se, CS₂Se, (NH₄)₂Se, (NH₄)HSe, BeSe, MgSe, CaSe,SrSe, PoSe and BaSe. A variety of chalcogenides are described inWO2005/041656, WO2005/041655 and WO2006/113914.

In certain embodiments, an active agent is an oxygen antagonist. Theterm “oxygen antagonist” refers to a substance that competes with oxygeninsofar as it used by a biological matter that requires oxygen for it tobe alive (“oxygen-utilizing biological matter”). Oxygen is typicallyused or needed for various cellular processes that create the biologicalmatter's primary source of readily utilizable energy. An oxygenantagonist effectively reduces or eliminates the amount of oxygen thatis available to the oxygen-utilizing biological matter, and/or theamount of oxygen that can be used by the oxygen-utilizing biologicalmatter. Thus, in some embodiments, an oxygen antagonist inhibits orreduces the amount of cellular respiration occurring in the cells, forinstance, by binding sites on cytochrome c oxidase that would otherwisebind to oxygen. One example of an oxygen antagonist is carbon monoxide.A variety of oxygen antagonists are described in WO2005/041656,WO2005/041655 and WO2006/113914.

In additional embodiments, an active agent is a stasis-inducingcompound, such as rotenone. Additional stasis-inducing compounds aredescribed, e.g., in US2010/0021387.

In certain embodiments, an active agent is an elemental reducing agent(ERA), such as iodide, sulfide or selenide.

In other embodiments, an active agent is a HIF modulating compound, avariety of which are described in US2009/0011051. The transcriptionfactor HIF (hypoxia inducible factor) system is a key regulator ofresponses to hypoxia, occupying a central position in oxygen homeostasisin a wide range of organisms. A large number of transcriptional targetshave been identified, with critical roles in angiogenesis,erythropoiesis, energy metabolism, inflammation, vasomotor function, andapoptotic/proliferative responses. The system is essential for normaldevelopment, and plays a key role in pathophysiological responses toischaemia/hypoxia. HIF is also important in cancer, in which it iscommonly upregulated, has major effects on tumor growth andangiogenesis. The HIF DNA binding complex consists of a heterodimer of αand β subunits. Regulation by oxygen occurs through hydroxylation of theα-subunits, typically of one or more proline residues, which are thenrapidly destroyed by the proteasome in oxygenated cells. This involvesbinding of HIFα-subunits by the von Hippel-Lindau tumor suppressorprotein (pVHL or VHL), with pVHL acting as the, or part of the,recognition component for a ubiquitin ligase that promotes ubiquitindependent proteolysis through interaction with a specific sequence orsequences in HIF α-subunits. In hypoxia, this process is suppressed,thereby stabilizing HIFα and permitting transcriptional activation viathe HIF α/β heterodimer. Certain compounds shown to affect metabolicactivity also act to stabilize HIF, thereby implicating such compoundsas preventative and treatment regimes for hypoxia, ischemia, stasis,and/or any other condition or state associated with HIF stabilization,such as hemorrhagic shock. Any of these compounds may be used accordingto the present invention.

In other embodiments, an active agent is a polychalcogenide.Representative polychalcogenides are described in WO2009/003061.

In further embodiments, an active agent is a polysulfide compound,examples of which are described in WO2010/045582.

Another active agent that can be used, e.g., in combination with ahalogen compound, is nitric oxide.

Certain compositions and methods of the present invention comprise achalcogenide, in combination with glutathione and/or a goitrogen. Asdiscussed herein, glutathione can stabilize chalcogenides in a reducedform, and goitrogens can inhibit production of thyroid hormone.

In certain embodiments, methods of the present invention includeproviding to a subject in need thereof any of the following agents orcompounds, alone or in combination, including but not limited to any ofthe combinations shown below:

a halogen compound;

a chalcogenide;

a goitrogen or other compound that reduces or inhibits thyroid hormoneproduction, activity or uptake;

a halogen compound and a chalcogenide;

a chalcogenide and glutathione;

a halogen compound and glutathione; and

a halogen compound, a chalcogenide, and glutathione.

In certain embodiments, a halogen compound is administered orcoformulated with an active agent used to treat CHF, e.g., CHF due toleft ventricular dysfunction, such as an angiotensin converting enzyme(ACE) inhibitor, a beta blocker, an aldosterone antagonist, anangiotension receptor blocker, or hydralazine with a nitrate. Inadditional embodiments, the another active agent is an anticoagulant, anantiplatelet therapy, an angiotensin II receptor blocker, a calciumchannel blocker, a diuretic, a vasodilator, or a statin.

Non-limiting examples of angiotensin converting enzyme inhibitors thatmay be used according to the present invention include: include:captopril (Capoten®), enalapril (Vasotec®), lisinopril (Prinivil®,Zestril®), benazepril hydrochloride (Lotensin®), fosinopril sodium(Monopril®), ramipril (Altace®), quinapril hydrochloride (Accupril®),perindopril erbumine (Aceon®), trandolapril (Mavik®), and moexiprilhydrochloride (Univasc®).

Non-limiting examples of nonselective beta blockers that may be usedaccording to the present invention include: alpurinol, bucindolol,carteolol, carvedilol (has additional α-blocking activity), labetalol(has additional α-blocking activity), nadolol, oxprenolol (has intrinsicsympathomimetic activity), penbutolol (has intrinsic sympathomimeticactivity), pindolol (has intrinsic sympathomimetic activity),propranolol, sotalol, timolol, and eucommia bark (herb).

Non-limiting examples of β₁-selective beta blockers that may be usedaccording to the present invention (also known as cardioselective)include: acebutolol (has intrinsic sympathomimetic activity), atenolol,betaxolol, bisoprolol, beliprolol, esmolol, metoprolol, and nebivolol(also increases nitric oxide release for vasodilation).

Non-limiting examples of β₂-selective agents that may be used accordingto the present invention include: butaxamine (weak α-adrenergic agonistactivity).

Non-limiting examples of beta blockers specifically indicated forchronic heart failure include: carvedilol, sustained-release metoprolol,bisoprolol, and nebivolol.

Non-limiting examples of angiotensin II blockers include: candesartan(Atacand®), eprosartan (Teveten®), irbesartan (Avapro®), losartan(Cozaar®), telmisartan (Micardis®) and valsartan (Diovan®).

In certain embodiments, methods of the present invention includeproviding to a subject in need, e.g., a subject diagnosed with or atrisk for chronic heart failure, thereof any of the following agents orcompounds, alone or in combination, including but not limited to any ofthe combinations shown below:

a halogen compound;

a halogen compound and glutathione;

a halogen compound and another active agent used to treat chronic heartfailure; and

a halogen compound, another active agent used to treat chronic heartfailure, and glutathione.

When used in combination, the two or more agents may be provided to thesubject in the same or in different compositions. In addition, whenthree or more agents are provided to a subject, two may be in onecomposition, and the third in a separate composition. The compositionsmay be administered concurrently, during overlapping time periods, orduring different time periods.

Compositions and Unit Dosage Forms

The present invention also includes compositions comprising a halogencompound, such as, e.g., an iodide or bromide. In particularembodiments, the composition further comprises one or more additionalactive agents, including any of those described herein. In certainembodiments, compositions of the present invention are pharmaceuticalcompositions. In particular embodiments, compositions of the presentinvention further comprise a pharmaceutically acceptable carrier,diluent, or excipient, such as, e.g., a buffer. In certain embodiments,compositions of the present invention are pharmaceutical compositionscomprising a halogen compound and glutathione. Particular compositionscomprise a halogen compound, e.g an iodide, and another compound used totreat or prevent chronic heart failure. Additional compositions of theinvention comprise glutathione, in combination with a chalcogenideand/or a halogen compound. In certain embodiments, a compositioncomprises glutathione, a chalcogenide, and a halogen compound. Inparticular embodiments, a composition of the invention comprisesglutathione and selenide. In particular embodiments, a composition ofthe invention comprises glutathione and sulfide. In certain embodiments,a composition of the present invention comprises iodide (or iodate),selenide and glutathione. In certain embodiments, a composition of thepresent invention comprises iodide (or iodate), sulfide and glutathione.In particular embodiments, at least a portion of the selenide or sulfideis in reduced form, and the glutathione inhibits the oxidation of thechalcogenide in the composition. The reduced form of chalcogenide may beany of those described herein. In particular embodiments, at least aportion of the iodide or iodate is in reduced form, and the glutathioneinhibits the oxidation of the halogen compound in the composition. Thereduced form of halogen compound may be any of those described herein.

In certain embodiments, the present invention includes compositionscomprising a halogen compound, e.g., a reduced form of a halogencompound. In related embodiments, the invention includes compositionscomprising a halogen compound, e.g., a reduced form of a halogencompound and one or more additional active agents, e.g., a chalcogenide,such as a reduced form of a chalcogenide, or an active agent used totreat chronic heart failure. In certain embodiments, the compositionfurther comprises glutathione. The invention further includes methodsthat involve providing to a subject a composition comprising a reducedform of a halogen compound, alone or in combination with providing tothe subject a composition comprising one or more additional activeagent, such as, e.g., a reduced form of a chalcogenide. In particularembodiments, a reduced form of a chalcogenide comprises a chalcogen,e.g., sulfur or selenium, in a −2 valence state. In particularembodiments, a single composition comprises both the halogen compoundand the one or more additional active agent or an active agent used totreat chronic heart failure. In certain embodiments of any of thecompositions of the invention, the halogen compound is a reduced form ofa halogen compound, which comprises a halogen in a −1 valence state,e.g., an iodide or bromide, such as sodium iodide. In certainembodiments of any of the compositions described herein, the compositionfurther comprises glutathione or another reducing agent.

In particular embodiments, the compositions are formulated to maintainthe halogen and/or chalcogenide in a reduced form when stored over aperiod of time. Thus, the compositions may be stable compositions ofreduced forms of halogen compounds and/or reduced forms ofchalcogenides, or salts or precursors thereof, whose effectiveness as atherapeutic may normally be compromised during manufacture and storage,as a result of oxidation reactions that produce oxidation products. Thecompositions of the present invention have increased shelf-life, areeasily and reproducibly manufactured, are designed for standard routesof administration, and are, therefore, advantageous in the treatment andprevention of a number of diseases, conditions and injuries. The presentinvention contemplates their use in methods of protecting biologicaltissue from disease or injury, particularly ischemic, hypoxic orreperfusion injury, as well as methods of treating or preventing injuryor disease in a subject.

In certain embodiments, a stable composition comprising a chalcogenideand/or a halogen compound comprises glutathione.

In additional embodiments, a composition comprises a goitrogen or anactive agent that inhibits or reduces thyroid hormone production,activity or uptake, alone or in combination with another active agent,such as a chalcogenide and/or halogen compound.

In certain embodiments of the compositions, a composition is consideredstable, i.e., a stable composition, if at least 90% of the halogencompound or the chalcogenide (if present in the composition), or boththe halogen compound and the chalcogenide (if both present in thecomposition), in the composition is present in reduced form for at leastone hour either when stored at room temperature, 4° C., 25° C., 40° C.or 50° C. In related embodiments, a composition is considered stable ifat least 70%, at least 80%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, or atleast 99% of the halogen compound, the chalcogenide, or both the halogencompound and the chalcogenide, in the composition is present in reducedform for at least one hour either when stored at room temperature orwhen stored at 4° C. In certain embodiments of the stable compositions,at least 90% of the halogen compound, the chalcogenide, or both thehalogen compound and the chalcogenide, in said composition is present insaid reduced form for at least one day, at least one week, at least onemonth, at least two months, at least four months, at least six months,or at least one year, either when stored at room temperature or whenstored at 4° C., 25° C., 40° C. or 50° C. In related embodiments, atleast 70%, at least 80%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, or atleast 99% of the halogen compound, the chalcogenide, or both the halogencompound and the chalcogenide, in the stable composition is present insaid reduced form for at least one day, at least one week, at least onemonth, at least two months, at least four months, at least six months,or at least one year, either when stored at room temperature or whenstored at 4° C. In particular embodiments, at least 98% of the halogencompound, the chalcogenide, or both the halogen compound and thechalcogenide, in the stable composition is present in said reduced formfor at least one month or at least six months when stored at 4° C. Inrelated embodiments, at least 70%, at least 80%, at least 91%, at least92%, at least 93%, at least 94%, at least 95%, at least 96%, at least97%, at least 98%, or at least 99% of the halogen compound, thechalcogenide, or both the halogen compound and the chalcogenide, in thestable composition is present in said reduced form for at least one day,at least one week, at least one month, at least two months, at leastfour months, at least six months, or at least one year, either whenstored at room temperature or when stored at room temperature or 25° C.In particular embodiments, at least 98% of the halogen compound, thechalcogenide, or both the halogen compound and the chalcogenide, in thestable composition is present in said reduced form for at least onemonth or at least six months when stored at room temperature or 25° C.In related embodiments, at least 70%, at least 80%, at least 91%, atleast 92%, at least 93%, at least 94%, at least 95%, at least 96%, atleast 97%, at least 98%, or at least 99% of the halogen compound, thechalcogenide, or both the halogen compound and the chalcogenide, in thestable composition is present in said reduced form for at least one day,at least one week, at least one month, at least two months, at leastfour months, at least six months, or at least one year, either whenstored at room temperature or when stored at 40° C. or 50° C. Inparticular embodiments, at least 98% of the halogen compound, thechalcogenide, or both the halogen compound and the chalcogenide, in thestable composition is present in said reduced form for at least onemonth or at least six months when stored at 40° C. or 50° C. In variousembodiments, the composition is a liquid pharmaceutical composition,while in other embodiments, the composition is a solid or powder, or isdried, lyophilized, or freeze-dried.

In particular embodiments, the present invention relates to a stableliquid composition comprising iodide, wherein the stable liquidcomposition comprises less than 1% of any of the following oxidationproducts of iodide (−1 oxidation state): hypoiodite (+1 oxidationstate), iodite (+3 oxidation state), iodate (+5 oxidation state), orperiodate (+7 oxidation state). In particular embodiments, the stableliquid composition comprising iodide comprises less than 1% iodine (I₂).

In certain embodiments, a reduced form of a halogen compound comprises ahalogen in a −1 valence state. In particular embodiments, a reduced formof a chalcogenide comprises a chalcogen e.g., sulfur or selenium, in a−2 valence state. In particular embodiments, the reduced form of ahalogen is a reduced form of the halogen, iodine, such as iodide. Inparticular embodiments, the reduced form of a halogen compound is NaI,KI, HI, CaI or AgI. In particular embodiments, the reduced form of achalcogenide is H₂Se, Na₂Se, NaHSe, HSe—, H₂S, NaHS, Na₂S, or HS—.

In particular embodiments, any of the compositions described hereincomprise a pharmaceutically acceptable carrier, diluent or excipient.Further, any of the compositions may comprise one or more of a buffer, areducing agent, a tonicity agent, a stabilizer, a surfactant, alycoprotectant, a polyol, an antioxidant, or a preservative. Inparticular embodiments, any of the composition described herein compriseglutathione.

In particular embodiments, compositions may comprise one or moresolvents. In particular embodiments, the solvent is water. In particularembodiments, the solvent is a phosphate-buffered saline.

Compositions of the present invention and methods of the presentinvention may include a halogen compound and/or a chalcogenide compound,or salt or precursor thereof, in any desired concentration. Theconcentration may be readily optimized, e.g., depending upon the type ofinjury or disease being treated and the route of administration, so asto deliver an effective amount in a convenient manner and over anappropriate time-frame.

In some embodiments, the concentration of halogen compound or salt orprecursor thereof present in a composition of the present invention isabout 0.0001 mM to about 100 M, about 0.0005 mM to about 50 M, about0.001 mM to about 10 M, about 0.001 mM to about 5 M, about 0.001 mM toabout 1 M, about 0.005 mM to about 10 M, about 0.005 mM to about 5 M,about 0.005 mM to about 1 M, about 0.005 mM to about 0.5 M, about 0.01mM to about 10 M, about 0.01 mM to about 5 M, about 0.01 mM to about 2M, about 0.1 mM to about 1 M, about 0.1 mM to about 0.5 M, about 0.5 mMto about 5 M, about 0.5 mM to about 2 M, about 0.5 mM to about 1 M,about 0.5 mM to about 0.5 M, about 1 mM to about 5 M, about 1 mM toabout 2 M, about 1 mM to about 1 M, about 1 mM to about 0.5 M, about 5mM to about 5 M, about 5 mM to about 2 M, about 5 mM to about 1 M, about5 mM to about 0.5 M, about 5 mM to about 0.25 M, about 10 mM to about 1M, about 10 mM to about 0.5 M, about 10 mM to about 0.25 M, or about 10mM, about 50 mM about 100 mM, or about 200 mM.

In some embodiments, the concentration of chalcogenide or chalcogenidecompound or salt or precursor thereof is about 0.001 mM to about 5,000mM, about 1 mM to about 1000 mM, about 10 mM to about 500 mM, about 50mM to about 500 mM, about 75 mM to about 250 mM, or about 95 mM to 150mM.

In particular embodiments, a composition comprises selenide at aconcentration of about 0.1 mM to about 1000 mM, about 1 mM to about 1000mM, about 5 mM to about 1000 mM, about 10 mM to about 1000 mM, about 10mM to about 750 mM, about 50 mM to about 500 mM, about 100 mM to about500 mM, about 10 mM to about 500 mM, 1 mM to about 500 mM, or about 10mM to about 250 mM.

In particular embodiments, a composition comprises sulfide at aconcentration of about 0.1 mM to about 1000 mM, about 1 mM to about 1000mM, about 5 mM to about 1000 mM, about 10 mM to about 1000 mM, about 10mM to about 750 mM, about 50 mM to about 500 mM, about 100 mM to about500 mM, about 10 mM to about 500 mM, 1 mM to about 500 mM, or about 10mM to about 250 mM.

In certain embodiments, the concentration of the reduced form of halogencompound and/or chalcogenide, e.g., selenide or sulfide, in acomposition of the present invention is about, at least about, or atmost about 0.001, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09,0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4,1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8,2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2,4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0 mM or M or more or any rangederivable therein (at standard temperature and pressure (STP)).

As used herein, the term “%” when used without qualification (as withw/v, v/v, or w/w) means % weight-in-volume for solutions of solids in s(w/v), % weight-in-volume for solutions of gases in s (w/v), %volume-in-volume for solutions of s in s (v/v) and weight-in-weight formixtures of solids and semisolids (w/w) (Remington's PharmaceuticalSciences (2005); 21st Edition, Troy, David B. Ed. Lippincott, Williamsand Wilkins).

In one embodiment, a composition comprises selenide or sulfide measuredat 80%-100% (w/v). In one embodiment, a composition comprises selenideor sulfide measured at 90%-100% (w/v). In one embodiment, a compositioncomprises selenide or sulfide measured at 95%-100% (w/v). In oneembodiment, a composition comprises sulfide measured at 98%-100% (w/v).

In certain embodiments, a composition comprises glutathione at aconcentration of about 1.5 μM to about 10 M, about 15 μM to about 1 M,about 150 μM to about 1 M, about 1.5 mM to about 1 M, about 10 mM toabout 500 mM, about 10 mM to about 250 mM, or about 100 mM, about 120mM, about 150 mM, about 170 mM, or about 200 mM.

In certain embodiments, a composition comprises a goitrogen or acompound that inhibits or impedes thyroid hormone production, activityor uptake at a concentration of about 0.1 μM to about 100 M, about 1.5μM to about 10 M, about 15 μM to about 1 M, about 150 μM to about 1 M,about 1 mM to about 1 M, about 10 mM to about 500 mM, about 10 mM toabout 250 mM, or about 100 mM, about 120 mM, about 150 mM, about 170 mM,or about 200 mM.

In certain embodiments, a composition of the invention comprises achalcogenide and glutathione, and optionally a halogen compound or saltor precursor thereof, wherein the concentration of the chalcogenide isabout 1 mM to about 1 M, about 10 mM to about 500 mM, or about 10 mM toabout 100 mM, the concentration of glutathione is about 100 μM to about1 M, about 1 mM to about 1 M, or about 10 mM to about 500 mM, and, ifpresent, the concentration of halogen compound is about 0.01 mM to about5 M, about 1 mM to about 0.5 M, or about 10 mM to about 250 mM. Incertain embodiments, the halogen compound is present in the composition.In particular embodiments, the halogen compound is an iodide or iodate,and the chalcogenide comprises selenium or sulfur. In certainembodiments, a composition of the invention comprises a halogen compoundor salt or precursor thereof and glutathione, and optionally achalcogenide, wherein the concentration of the halogen compound is about0.01 mM to about 5 M, about 1 mM to about 0.5 M, or about 10 mM to about250 mM, the concentration of glutathione is about 100 μM to about 1 M,about 1 mM to about 1 M, or about 10 mM to about 500 mM, and, ifpresent, the concentration of halogen compound is about about 1 mM toabout 1 M, about 10 mM to about 500 mM, or about 10 mM to about 100 mM.In certain embodiments, the chalcogenide is present in the composition.In particular embodiments, the halogen compound is an iodide or iodate,and the chalcogenide comprises selenium or sulfur. In particularembodiments of any of these compositions, the composition is formulatedfor oral delivery, or is an oral dosage form, the halogen compound (whenpresent) comprises iodine (e.g., iodide or iodate), and the chalcogenide(when present) comprises selenium. In certain embodiments, thechalcogenide is selenide or selenide. In particular embodiments, thecomposition is formulated for intravenous administration, and thehalogen compound (if present) is iodide or iodate, and the chalcogenide(if present) is in a reduced form, e.g., selenide or sulfide. In oneembodiment, the composition comprises selenide, iodide and glutathione,each within any of the concentration ranges or at a concentrationdescribed herein.

In particular embodiments, the pH of a composition of the presentinvention is in the range of (3.0-12.0), while in other embodiments, thepH is in the range of (5.0-9.0). The pH of the pharmaceuticalcomposition may be adjusted to a physiologically compatible range. Forexample, in one embodiment, the pH of the stable composition is in therange of 6.5-8.5. In other embodiments, the compositions of the presentinvention have a pH in the range of 7.5-8.5 or 7.4-9.0.

In particular embodiments, oxygen is present in a composition of thepresent invention at a concentration in the range of 0 μM⁻⁵ μM or 0 μM⁻¹μM or 0 μM^(−0.1) μM or 0 μM-0.01 μM. In particular embodiments, oxygenis present in the composition at a concentration of less than 3 μM, lessthan 1 μM, less than 0.1 μM, less than 0.01 μM, or less than 0.001 μM.

In certain embodiments, the compositions of the present invention mayfurther comprise a limited amount of oxidation products. Oxidationproducts that may be present in various embodiments of the presentinvention include, but are not limited to, iodine, iodate, bromine,bromate, selenite, thioselenate, polyselenides, elemental selenium,selenate, sulfite, sulfate, thiosulfate, polysulfides, dithionate,polythionate, and elemental sulfur. In various embodiments, one or moreof these oxidation products is present in a composition in an amountless than 10%, less than 5.0%, less than 2.0%, less than 1.0%, less than0.5%, less than 0.2%, less than 0.1%, less than 0.05%, or less than0.01% (w/v) of the total halogen compound and/or chalcogenide in thecomposition.

In one embodiment, a composition has an osmolarity in the range of200-400 mOsmol/L. NaCl may be used as an excipient to adjust osmolality.

In certain embodiments, isotonicity of the compositions is desirable asit results in reduced pain upon administration and minimizes potentialhemolytic effects associated with hypertonic or hypotonic compositions.Thus, the compositions of the invention not only have increased storagestability, but also have the added benefit of substantially reduced painupon administration when compared with formulations using other moretraditional buffer systems consisting of an acid and a salt form of theacid.

Stable liquid compositions comprising reduced forms of chalcogenides,including sulfide or selenide, and methods of manufacturing the same,have been described, e.g., in PCT Application Publication No.WO2008/043081, U.S. Provisional Application No. 61/659,311, and PCTApplication Publication No. WO2013/188528. Any of these composition maybe used to treat or prevent any of the diseases, disorders or injuriesdescribed herein

Compositions of the invention comprising a reduced form of achalcogenide and further comprising glutathione may be prepared by oneof skill in the art in a similar manner, e.g., by preparing acomposition comprising the reduced form of chalcogenide and adding theglutathione. Similarly, compositions comprising a reduced form of achalcogenide and a reduced form of a halogen compound may be prepared byone of skill in the art in a similar manner, e.g., by preparing acomposition comprising the reduced form of chalcogenide and combining itwith a reduced form of a halogen compound or with a compositioncomprising a reduced form of a halogen compound.

WO2008/043081 describes compositions comprising a stable liquidpharmaceutical chalcogenide or chalcogenide compound or salt orprecursor thereof in a pharmaceutically acceptable carrier, wherein theconcentration, pH and oxidation products of said chalcogenide orchalcogenide compound or salt remain within a range of acceptancecriteria after storage of said liquid pharmaceutical composition. Invarious embodiments, the chalcogenide compound or chalcogenide salt isselected from the group consisting of: H₂S, Na₂S, NaHS, K₂S, KHS, Rb₂S,CS₂S, (NH₄)₂S, (NH₄)HS, BeS, MgS, CaS, SrS, and BaS. In otherembodiments, the chalcogenide compound or chalcogenide salt is selectedfrom the group consisting of: H₂Se, Na₂Se, NaHSe, K₂Se, KHSe, Rb₂Se,CS₂Se, (NH₄)₂Se, (NH₄)HSe, BeSe, MgSe, CaSe, SrSe, PoSe and BaSe.According to the present invention, said compositions may furthercomprise a halogen compound, e.g., iodide, glutathione, or a goitrogen.

In particular embodiments of stable liquid or aqueous compositions, thechalcogenide compound or chalcogenide salt is sulfide and has aconcentration in the range of 95 mM to 150 mM.

In particular embodiments wherein said chalcogenide compound orchalcogenide salt is sulfide, said sulfide is present in amounts rangingfrom about 80% to about 100%, about 90% to 100%, or about 95% to 100% byw/v.

In particular embodiments, the liquid is sodium hydroxide.

In certain embodiments, the composition has a pH in the range of 6.5 to8.5. In one embodiment, the composition has an oxygen content of lessthan or equal to 5 μM.

In one embodiment, the composition further comprises one or moreoxidation products selected from polysulfide, sulfite, sulfate andthiosulfate. The oxidation products may be sulfate in the range of(0%-1.0%), or sulfite in the range of (0%-1.0%), or polysulfide in therange of (0%-1%) or thiosulfate in the range of (0%—1.0%).

The storage period may be about 3 months at a range of(23°−27°) or 6months at a range of (23°−27°). In one embodiment, the composition hasan osmolarity in the range of 250-330 mOsmol/L. It may be isotonic ornear isotonic.

In certain embodiments, a composition is a sterile, stable aqueouspharmaceutical composition comprising a solution of deoxygenated waterand sodium sulfide nonahydrate, wherein the sodium sulfide nonahydrateproduces Na₂S, H₂S and HS⁻, wherein the concentration of H₂S is in therange of 1.0 mg/mL to 4.0 mg/mL; wherein the solution is adjusted to apH in the range of 7.5 to 8.5 by adding HCl to the solution; wherein thesolution has an osmolarity in the range of 250-330 mOsmol/L; wherein thesolution further comprises sulfide oxidation products selected from thegroup consisting of polysulfide, sulfite, sulfate and thiosulfate;wherein the oxidation products comprise sulfate in the range of(0%-1.0%), sulfite in the range of (0%-1.0%), polysulfide in the rangeof (0%-1%) or thiosulfate in the range of (0%-1.0%); wherein thesolution is isotonic or near isotonic; and, wherein the solution isstable for at least four months. In particular embodiments, thecomposition has an oxygen content in the range of 0 μM to 5 μM, 0 μM to3 μM, 0.01 μM to 1 μM, or 0.001 μM to 1 μM. In certain embodiments, thecomposition further comprises a halogen compound, such as, e.g., aniodide or a bromide, such as sodium iodide. Such a composition mayfurther comprise glutathione or a goitrogen or other agent that impedesor inhibits thyroid hormone production, activity or uptake.

In certain embodiments, the composition is a sterile, stablepharmaceutical composition comprising: a solution of deoxygenated waterand Na₂S, wherein the Na₂S produces HS⁻, wherein the solution has a pHin the range of 7.8 to 8.2; wherein the solution has an osmolarity inthe range of 250-330 mOsmol/L; wherein the solution further comprisessulfide oxidation products selected from the group consisting ofpolysulfide, sulfite, sulfate and thiosulfate; wherein the oxidationproducts comprise sulfate in the range of (0%-1.0%), sulfite in therange of (0%-1.0%), polysulfide in the range of (0%-1%) or thiosulfatein the range of (0%-1.0%); wherein the solution is stable for at leastfour months; and, wherein the solution is isotonic or near isotonic. Incertain embodiments, the composition further comprises a halogencompound, such as, e.g., an iodide or a bromide, such as sodium iodide.Such a composition may further comprise glutathione (or another compoundthat inhibits or impedes thyroid hormone production or activity or agoitrogen).

In certain embodiments, the composition is a sterile, stable, liquidpharmaceutical composition made by the steps, in any suitable order,comprising: making a solution of Na₂S by rinsing Na₂S.9H₂O crystals withdeoxygenated, distilled and deionized water; bubbling N₂, a mixture ofN₂/CO₂, or a mixture of N₂/H₂S through the solution generating the Na₂Sthat produces HS, and, adjusting the pH of the solution to a pH in therange of 7.5 to 8.5 by adding HCl to the solution; wherein said solutionhas an osmolarity in the range of 250-330 mOsmol/L; wherein saidsolution further comprises sulfide oxidation products selected from thegroup consisting of polysulfide, sulfite, sulfate and thiosulfate;wherein the oxidation products comprise sulfate in the range of(0%-1.0%), sulfite in the range of (0%-1.0%), polysulfide in the rangeof (0%-1%) or thiosulfate in the range of (0%-1.0%); and, wherein saidcomposition is isotonic or near isotonic. In certain embodiments, thecomposition further comprises a halogen compound, such as, e.g., aniodide or a bromide, such as sodium iodide. Such a composition mayfurther comprise glutathione or a goitrogen or other agent that impedesor inhibits thyroid hormone production, activity or uptake.

In various embodiments of the above compositions comprising achalcogenide, the concentration of HS⁻ is in the range of 1 mM to 250mM, 10 mM to 200 mM or 95 mM to 150 mM. In one embodiment of the abovecompositions comprising a chalcogenide, the concentration of HS⁻ is inthe range of 1 mM to 250 mM, 10 mM to 200 mM or 95 mM to 150 mM. In oneembodiment of the above compositions comprising a chalcogenide, theconcentration of HS⁻ is in the range of 1 mM to 250 mM, 10 mM to 200 mMor 95 mM to 150 mM.

The present invention further includes kits comprising composition(s) ofthe present invention. In certain embodiments, such kits comprise one ormore containers to store the composition(s) of the present invention. Inone embodiment, a composition is stored in the container under an inertor noble gas, and the container is a sealed and has an oxygenimpermeable light-protective container (e.g., an amber vial). In certainembodiments, a kit comprises a composition comprising a halogencompound, e.g., a reduced form of iodine, such as iodided, and acomposition comprising an active agent used to treat chronic heartfailure.

In certain embodiments, a composition is packaged in an impermeablecontainer. “Impermeable container” refers to containers that provide abarrier to the passage of gas molecules. Impermeable containers areknown to those skilled in the art and include, but are not limited to,“i.v. bags” or syringes comprising a gas impermeable constructionmaterial, or a sealed glass vial. In particular embodiments, thecomposition may be packaged into an impermeable container containing aninert atmosphere, an inert gas, or a noble gas. Noble gas refers tohelium (He), neon (Ne), argon (Ar), krypton (Kr), xenon (Xe), and radon(Rn). Inert gas refers to nitrogen (N2). The term “inert atmosphere”refers to a nitrogen or argon atmosphere in a container. In particularembodiments, the container comprises a reduced oxygen or oxygen-freeenvironment. A “reduced oxygen environment” is an environment having anoxygen concentration of less than 100 parts per million. The compositionmay be packaged in a light-protective vial or container, e.g., ambervials. In one embodiment, the composition is sealed and stored in aglass ampoule.

In some embodiments, compositions of the present invention comprise oneor more excipients included to prevent oxidation of the halogen compoundand/or chalcogenide during storage, where storage is in the range of oneto twelve months or longer. In some embodiments, storage is in the rangeof one to six months. In some embodiments, storage is in the range ofthree to six months. In some embodiments, storage is in the range offour to five months. Embodiments of the present invention may use asingle excipient or a combination of excipients. There are many suitableexcipients. Examples include chelators, pH modifying agents, reducingagents, antioxidants, spin-trap agents and preservatives.

In one embodiment, compositions of the present invention may optionallycontain chelators or chelating agents. Chelating agents yielding solublemetal complexes are also called sequestering agents. A chelating agenttypically has at least two functional groups that donate a pair ofelectrons to the metal, such as —O—, —NH₂ or —COO—. Examples ofnaturally-occurring chelators include carbohydrates, includingpolysaccharides, organic acids with more than one coordination group,lipids, steroids, amino acids and related compounds, peptides,phosphates, nucleotides, tetrapyrrole, ferrioxamines, ionophores, suchas gramicidin, monensin, valinomycin, and phenolics. Examples ofsynthetic chelators include, but are not limited to,Diethylenetriaminepentaacetic acid (DTPA), Diethylenetriaminepentaaceticacid pentasodium salt (DTPA5), CaDTPAH, dimercaprol (BAL), deferoxamine,desferal, 2,2′-Bipyridyl DimercaptopropanolEthylenediaminotetraaceticacid, Ethylenedioxy-diethylene-dinitrilo-tetraacetic acid (EDTA),CaNa2ethylenediaminetetraacetic acid, Ethyleneglycol-bis-(2-aminoethyl)-N,N,N′,N′-tetraacetic acid (EGTA), ionophores,Nitrilotriacetic acid (NTA), ortho-Phenanthroline, Salicylic acid,succimer (meso-2,3-dimercaptosuccinic acid, (DMSA), Triethanolamine(TEA), N-(2-Hydroxyethyl)ethylenediamine-N,N′,N′-triacetic acidTrisodium salt (HEDTA), Nitrilotriacetic acid (NTA). In one embodiment,the synthetic chelator is DTPA. In certain embodiments, theconcentration of DTPA is about, at least about, or at most about 0,0.001, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2,0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0 mM or M or any range derivabletherein. In one embodiment, the DTPA is in the range of 0.1 mM to 50 mM.In one embodiment, the synthetic chelator consists of DTPA5. In certainembodiments, the concentration of DTPA5 is in the range of(0.0001%-0.1%) (w/v). In another embodiment, DTPA5 is in the range of(0%-1.0%) (w/v). In one embodiment, DTPA5 is in the range of (0% to0.01%) (w/v). In one embodiment, the synthetic chelator is CaDTPA. Incertain embodiments, the concentration of CaDTPA is in the range of(0.0001%-0.1%) (w/v). In one embodiment, CaDTPA is in the range of (0%to 0.01%) (w/v). In another embodiment, CaDTPA is in the range of(0%-1.0%) (w/v). In one embodiment, the synthetic chelator isdeferoxamine. In certain embodiments, the concentration of deferoxamineis about, at least about, or at most about 0, 0.001, 0.01, 0.02, 0.03,0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7,0.8, 0.9, 1.0 mM or M, or any range derivable therein. In oneembodiment, the deferoxamine is in the range of 0.1 mM to 10 mM. In oneembodiment, the synthetic chelator is EDTA. In certain embodiments, theconcentration of EDTA is about, at least about, or at most about 0,0.001, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2,0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0 mM or M, or any range derivabletherein. In a certain embodiment, EDTA is in the range of 0%-1% (w/v).In another embodiment, EDTA is in the range of 0.0001%-0.1% (w/v). Inanother embodiment, EDTA is in the range of 0%-1.0% (w/v). In oneembodiment, EDTA is in the range of 0% to 0.01% (w/v).

Compositions of the present invention may further comprise one or morepH modifying agents. pH modifying agents, include, but are not limitedto, inorganic salts, such as zinc carbonate, magnesium carbonate,calcium carbonate, magnesium hydroxide, calcium hydrogen phosphate,calcium acetate, calcium hydroxide, calcium lactate, calcium maleate,calcium oleate, calcium oxalate, calcium phosphate, magnesium acetate,magnesium hydrogen phosphate, magnesium phosphate, magnesium lactate,magnesium maleate, magnesium oleate, magnesium oxalate, sodium chloride,sodium carbonate, sodium bicarbonate, potassium hydroxide, potassiumphosphate, sodium bicarbonate, thioglycolic acid, zinc acetate, zinchydrogen phosphate, zinc phosphate, zinc lactate, zinc maleate, zincoleate, zinc oxalate, and combinations thereof. Other pH modifyingagents include, e.g., acetic acid, fumaric acid, malic acid, nitricacid, phosphoric acid, propionic acid, sulfuric acid, tartaric acid,carbon dioxide, carbonic acid, N-methyl-D-glucamine,4-(2-hydroxyethyl)-morpholine, Tromethamine, Orotic acid, andhydrochloric acid. In one embodiment, the pH modifying agent is sodiumhydroxide.

A pH modifying agent may serve as a buffering agent when it is added toan already acidic or basic solution, which it then modifies andmaintains at a new pH (see: The United States Pharmacopeia-NationalFormulary 29th Edition, (2006) Rockville, Md.; Stahl, P. Wermuth, C. ed.Handbook of Pharmaceutical Salts Properties, Selection and Use. Wiley(2002)).

In certain embodiments, compositions of the present invention includeone more excipients that are reducing agents, such as, e.g., glutathione(see: U.S. Pat. No. 6,586,404), tris(2-carboxyethyl)phosphinehydrochloride (TSEP), I-cysteine, cysteine or methionine. In oneembodiment, the reducing agent is glutathione (see: Vincent et al.,Endocrine Reviews (2004) 25:612-628), dithiothreitol (DTT) (Weir et al.,Respir and Physiol Biol; (2002) 132:121-30) or dithioerythritol (DTE).In certain embodiments, the concentration of glutathione is about, atleast about, or at most about 0, 0.001, 0.01, 0.02, 0.03, 0.04, 0.05,0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0mM or M or more or any range derivable therein. In certain embodiments,the concentration of dithiothreitol (DTT), which present at about, atleast about, or at most about 0, 0.001, 0.01, 0.02, 0.03, 0.04, 0.05,0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0mM or 1 M, or any range derivable therein. In certain embodiments, thereducing agent is dithioerythritol (DTE), is about, at least about, orat most about 0, 0.001, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08,0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0 mM or M, or anyrange derivable therein.

Compositions of the present invention may optionally comprise a freeradical scavenger or antioxidant. Examples of free radical scavengers orantioxidants include, but are not limited to, ascorbic acid (vitamin C),D-alpha tocopherol acetate, DL-alpha-tocopherol (vitamin E), melatonin,sodium bisulfite, sodium sulfite, sodium metabisulfite, Trolox(6-hydroxy-2,5,7,8-tetramethyl chroman-2-carboxylic acid),Tris(2-Carboxyethyl)phosphine Hydrochloride (TCEP), melatonin,dithionite, pyrosulfite, cysteine, potassium disulfite, sodiumthioglycolate, thioethylene glycol, L-threoascobic acid, acetylsalicylicacid, salicylic acid, lecithin, ascorbyl palmitate, butylatedhydroxyanidole, ascorbic acid, butylated hydroxyanisole, butylatedhydroxyquinone, butylhydroxyanisol, hydroxycomarin, butylatedhydroxytoluene, cephalm, ethyl gallate, propyl gallate, octyl gallate,lauryl gallate, propylhydroxybenzoate, trihydroxybutylrophenone,dimethylphenol, lecithin, ethanolamine, meglumine and combinationsthereof (see US 2005/0106214). In one embodiment, the antioxidant agentis a spin-trap agent. Examples of spin-trap agents include, but are notlimited to, N-t-butyl-phenylnitrone (PBN) (see: Kotake, Y., AntioxidRedox Signal (1999) 481), 4-Hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl(TEMPOL) (Gariboldi, M. B., et al. (2000), Free Radic. Biol. Med.29:633; Miura, Y., et al. J. Radiat. Res. (Tokyo) (2000) 41:103;Mota-Filipe, H., et al. (1999), Shock 12:255R: 22-41; S: 39-262,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO) (see: Lapchak, et al.,Stroke (2001) 32:147-53);(disodium-[(tert-butylimino)methyl]benzene-1,3-disulfonate N-oxide(NXY-059) (see: Lapchak et al., CNS Drug Rev (2003) 9:253-62). In someembodiments, the spin-trap agent is TEMPO, which is present in the rangeof 0 mg/kg-1,000 mg/kg. In some embodiments, the spin-trap agent isTEMPO and is present in the range of 100 mg/kg-1,000 mg/kg. In anotherembodiment, the spin-trap agent is TEMPO and is present in the range of0 mg/kg-100 mg/kg.

Composition of the present invention may optionally comprise apreservative. As used herein, the term “preservative” is intended tomean a compound used to prevent the growth of microorganisms. Suchcompounds by way of example and without limitation, include benzalkoniumchloride, benzethonium chloride, benzoic acid, benzyl alcohol, butylatedhydroxyanisole (BHA), cetrimonium bromide, cetylpyridinium chloride,chlorobutanol, chlorocresol, cresol, methylparaben sodium, phenol,pheenoxyethanol, phenylethyl alcohol, phenylmercuric acetate,phenylmercuric nitrate, phenylmercuric acetate, thimerosal, metacresol,myristylgamma picolinium chloride, potassium benzoate, potassiumsorbate, sodium benzoate, sodium propionate, sorbic acid, thioglycerol,thimerosal, thymol, and methyl, ethyl, propyl or butyl parabens andothers known to those of ordinary skill in the art. Such preservativesare used in stable compositions at typical concentrations in accordancewith acceptable pharmaceutical practices, such as described. (see: TheUnited States Pharmacopeia—National Formulary 29th Edition, (2006)Rockville, Md.; Remington's Pharmaceutical Sciences (2005) 21st Edition,Troy, D B, Ed. Lippincott, Williams and Wilkins). In a certainembodiment, the preservative is benzyl alcohol and is present in therange of 0%-1.0% (w/v). In one embodiment, the preservative is benzylalcohol and is present in the range of 0%-0.5% (w/v). In one embodiment,the preservative is phenol in the range of 0%-0.5% (w/v). In a certainembodiment, the preservative is methyl paraben in the range of(0.0%-0.25% (w/v). In a certain embodiment, the preservative is ethylparaben in the range of 0%-0.25% (w/v). In a certain embodiment, thepreservative is propyl paraben in the range of 0%-0.25% (w/v). In acertain embodiment, the preservative is butyl paraben, in the range of0%-0.4% (w/v). In a certain embodiment, the preservative is benzalkoniumchloride in the range of 0%-0.02% (w/v).

In particular embodiments, any of the compositions of the presentinvention comprise may comprise both a reduced form of a halogencompound and a reduced form of a chalcogenide. In addition, acomposition may comprise two or more halogen compounds and/or two ormore chalcogenides.

The present invention also includes unit dosage forms of compositions ofthe present invention. In certain embodiments, the unit dosage formcomprises or consists of an effective amount of a halogen compound fortreating or preventing a disease, condition, or injury, including any ofthose described herein, in a subject in need thereof. In certainembodiments, the unit dosage form comprises or consists of an effectiveamount of a chalcogenide for treating or preventing a disease,condition, or injury, including any of those described herein, in asubject in need thereof. In certain embodiments, a unit dosage formfurther comprises glutathione in an amount effective to maintain thechalcogenide and/or halogen compound in a reduced form under any of theconditions described herein. In particular embodiments, the unit dosageform is formulated for intravenous administration, administration byinfusion, or oral administration. In certain embodiments, as usedherein, the term “prevention” includes inhibiting or impeding the onsetor progression of a disease or injury, or reducing the amount of injuryor damage caused by a disease or injury.

In particular embodiments, a unit dosage form comprising a halogencompound, such as an iodide or NaI, comprises or consists of about 0.005mg to about 5000 mg, about 0.05 to about 1000 mg, about 0.5 mg to about100 mg, about 1 mg to about 100 mg, about 2.5 mg to about 100 mg, about0.5 mg to about 50 mg, about 1 mg to about 50 mg, about 2.5 mg to about50 mg, about 5 mg to about 50 mg, about 10 mg to about 50 mg, or about 1mg, about 2 mg, about 5 mg, about 10 mg, or about 15 mg. In relatedembodiments, the unit dosage form comprises less than or equal to 150mg, less than or equal to 125 mg, less than or equal to 100 mg, lessthan or equal to 75 mg, less than or equal to 50 mg, less than or equalto 25 mg, or less than or equal to 10 mg of the halogen compound. Incertain embodiments, the unit dosage form comprises between about 1 mgand about 150 mg (including any interval in this range), between about 1mg and about 125 mg, between about 1 mg and about 100 mg, between about1 mg and about 75 mg, between about 1 mg and about 50 mg, between about1 mg and about 25 mg or between about 1 mg and about 10 mg of thehalogen compound. In certain embodiments, the unit dosage form comprisesabout 150 mg, about 125 mg, about 100 mg, about 75 mg, about 50 mg,about 25 mg or about about 10 mg of the halogen compound. In certainembodiments, the unit dosage form comprises less than or equal to 1000mg, less than or equal to 800 mg, less than or equal to 700 mg, lessthan or equal to 500 mg, less than or equal to 250 mg, less than orequal to 200 mg, or less than or equal to 150 mg of the halogencompound. In certain embodiments, the unit dosage form comprises betweenabout 100 mg and about 1000 mg (including any interval in this range),between about 150 mg and about 800 mg, between about 200 mg and about700 mg, between about 250 mg and about 600 mg, between about 300 mg andabout 500 mg, between about 350 mg and about 450 mg or between about 300mg and about 700 mg of the halogen compound. In certain embodiments, theunit dosage form comprises about 200 mg, about 300 mg, about 400 mg,about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, orabout about 1000 mg of the halogen compound.

In some embodiments, including, e.g., embodiments where the unit dosageform is formulated as a liquid, e.g., for intravenous administration oradministration by infusion, the concentration of halogen compound orsalt or precursor thereof present in a unit dosage form of the presentinvention is about 0.0001 mM to about 100 M, about 0.0005 mM to about 50M, about 0.001 mM to about 10 M, about 0.001 mM to about 5 M, about0.001 mM to about 1 M, about 0.005 mM to about 10 M, about 0.005 mM toabout 5 M, about 0.005 mM to about 1 M about 0.005 mM to about 0.5 M,about 0.01 mM to about 10 M, about 0.01 mM to about 5 M, about 0.01 mMto about 2 M, about 0.1 mM to about 1 M, about 0.1 mM to about 0.5 M,about 0.5 mM to about 5 M, about 0.5 mM to about 2 M, about 0.5 mM toabout 1 M, about 0.5 mM to about 0.5 M, about 1 mM to about 5 M, about 1mM to about 2 M, about 1 mM to about 1 M, about 1 mM to about 0.5 M,about 5 mM to about 5 M, about 5 mM to about 2 M about 5 mM to about 1M, about 5 mM to about 0.5 M, about 5 mM to about 0.25 M, about 10 mM toabout 1 M, about 10 mM to about 0.5 M, about 10 mM to about 0.25 M, orabout 10 mM, about 50 mM about 100 mM, or about 200 mM. The unit dosageform may further comprise one or more pharmaceutically acceptablediluents, excipients or carriers.

In certain embodiment, the unit dosage form comprises iodine, e.g., NaI,and the effective amount is greater than or equal to about 150 μg,greater than or equal to about 300 μg, greater than or equal to about500 μg, greater than or equal to about 1 mg, greater than or equal toabout 2 mg, greater than or equal to about 5 mg, greater than or equalto about 10 mg, greater than or equal to about 15 mg, or greater than orequal to about 20 mg. In certain embodiments, the effective amount is150 μg to 1000 mg, 300 μg to 1000 mg, 500 μg to 1000 mg, 1 mg to 1000mg, 2 mg to 1000 mg, 5 mg to 1000 mg, 10 mg to 1000 mg, 150 μg to 100mg, 300 μg to 100 mg, 500 μg to 100 mg, 1 mg to 100 mg, 2 mg to 100 mg,5 mg to 100 mg, or 10 mg to 100 mg. In certain embodiments, theeffective amount is 150 μg to 50 mg, 300 μg to 20 mg, 500 μg to 10 mg, 1mg to 20 mg, 1 mg to 10 mg, or about 5 mg, about 10 mg, about 15 mg, orabout 20 mg. In other embodiments, the effective amount is between about1 mg and about 150 mg (including any interval in this range), betweenabout 1 mg and about 125 mg, between about 1 mg and about 100 mg,between about 1 mg and about 75 mg, between about 1 mg and about 50 mg,between about 1 mg and about 25 mg or between about 1 mg and about 10 mgof the halogen compound. In certain embodiments, the effective amount isabout 150 mg, about 125 mg, about 100 mg, about 75 mg, about 50 mg,about 25 mg or about about 10 mg of the halogen compound. In certainembodiments, the effective amount comprises less than or equal to 1000mg, less than or equal to 800 mg, less than or equal to 700 mg, lessthan or equal to 500 mg, less than or equal to 250 mg, less than orequal to 200 mg, or less than or equal to 150 mg of the halogencompound. In certain embodiments, the effective amount is between about100 mg and about 1000 mg (including any interval in this range), betweenabout 150 mg and about 800 mg, between about 200 mg and about 700 mg,between about 250 mg and about 600 mg, between about 300 mg and about500 mg, between about 350 mg and about 450 mg or between about 300 mgand about 700 mg of the halogen compound. In certain embodiments, theeffective amount is about 200 mg, about 300 mg, about 400 mg, about 500mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, or aboutabout 1000 mg of the halogen compound. In particular embodiments, theeffective amount is the amount per day.

In certain embodiments, a unit dosage form comprises or consists of aneffective amount of a chalcogenide for treating or preventing a disease,condition, or injury, including any of those described herein, in asubject in need thereof. In particular embodiments, a unit dosage formof a chalcogenide comprises about 0.005 mg to about 5000 mg, about 0.05mg to about 5000 mg, about 0.1 mg to about 5000 mg, about 0.005 mg toabout 2000 mg, about 0.05 mg to about 2000 mg, about 0.1 mg to about2000 mg, about 0.5 mg to about 2000 mg, about 1 mg to about 1000 mg,about 2 mg to about 500 mg, about 5 mg to about 500 mg, about 10 mg toabout 100 mg, about 10 mg, about 20 mg, about 50 mg, about 60 mg, about70 mg, about 80 mg, about 100 mg, or about 150 mg.

In certain embodiments, the unit dosage form comprises or consists of aneffective amount of both a halogen compound and a chalcogenide, whereinthe halogen compound and chalcogenide are present in the unit dosageform in an amount useful for treating or preventing a disease,condition, or injury, including any of those described herein, in asubject in need thereof. It is understood that the effective amount ofeach of the halogen compound and chalcogenide may be different when theyare used in combination as compared to when they are used individually.

In certain embodiments, the unit dosage form comprises or consists of aneffective amount of both a halogen compound and an active agent used totreat chronic heart failure, wherein the halogen compound and the activeagent are present in the unit dosage form in an amount useful fortreating or preventing a disease, condition, or injury, such as chronicheart failure, in a subject in need thereof. It is understood that theeffective amount of each of the halogen compound and chalcogenide may bedifferent when they are used in combination as compared to when they areused individually.

In certain embodiments, the unit dosage form comprises or consists of aneffective amount of a composition comprising both a chalcogenide andglutathione, a composition comprising both a halogen compound andglutathione, or a composition comprising all of a halogen compound, achalcogenide, and glutathione, wherein the halogen compound (if present)and chalcogenide (if present) are present in the composition in anamount useful for treating or preventing a disease, condition, orinjury, including any of those described herein, in a subject in needthereof. The glutathione may be present in an amount sufficient toinhibit oxidation of the chalcogenide and/or halogen compound. It isunderstood that the effective amount of each of the halogen compound andchalcogenide may be different when they are used in combination ascompared to when they are used individually. In certain embodiments, thecomposition comprises about 15 mM to about 500 mM glutathione or about10 mM to about 500 mM glutathione.

In certain embodiments, a composition of the invention may be formulatedin a dosage form suitable for oral or parenteral administration. Inaddition, in particular embodiments, a composition of the invention maybe in the form of an immediate or modified release formulation. Forexample, formulations of the halogen compound and/or chalcogenide can beused to provide controlled release, in which the release of thecompound(s) is controlled and regulated to allow less frequency ofdosing or to improve the pharmacokinetic or toxicity profile of a givenactive agent.

In general the amount of the active compound present in a composition orunit dosage form depends inter alia on the specific compound andformulation, the age and condition of the subject, and the injury,condition, or disease being treated or prevented, the route ofadministration and the dosage frequency.

The dosage frequency also depends on the injury, condition or diseasebeing treated or prevented, the amount or concentration of the compound,the specific composition used, the route of administration and mayincorporate subject-specific variation including, but not limited toage, weight, gender, or overall health. For example, an oral compositionmay be administered prior to surgery to prevent ischemia/reperfusioninjury during or post-surgery, in which case it may be administered oneor more times prior to surgery, optionally in either an immediate ormodified or controlled release formulation. In other situations, forexample, an oral or intravenous composition may be administeredfollowing injury or the occurrence of a medical condition (e.g., strokeor heart attack), in which case it may be advantageous to use animmediate release formulation to achieve a relatively fast onset oftreatment or prevention of ischemia/reperfusion injury at the site ofinjury or medical condition.

In certain embodiments, a unit dosage form suitable for oraladministration is in the form of a pill, drenches (aqueous ornon-aqueous solutions or suspensions), boluses, powders, granules,polymer release formulations, pastes for application to the tonguetablet, caplet or a capsule. A pill is a small, round, solidpharmaceutical oral dosage form that was in use before the advent oftablets and capsules. In colloquial usage, tablets, capsules, andcaplets are still often referred to as “pills” collectively. In certainembodiments, pills are made by mixing the active ingredients with anexcipient such as glucose syrup in a mortar and pestle to form a paste,then divided into suitable sizes, and often coated with sugar to makethem more palatable.

A tablet is a pharmaceutical dosage form that comprises a mixture ofactive substances and excipients, usually in powder form, pressed orcompacted from a powder into a solid dose. The excipients can includediluents, binders or granulating agents, glidants (flow aids) andlubricants to ensure efficient tabletting; disintegrants to promotetablet break-up in the digestive tract; sweeteners or flavours toenhance taste; and pigments to make the tablets visually attractive,etc. A polymer coating is often applied to make the tablet smoother andeasier to swallow, to control the release rate of the active ingredient,or to make it more stable. Sizes of tablets to be swallowed typicallyrange from a few millimeters to about a centimeter. Some tablets are inthe shape of capsules, and are called “caplets”.

Capsules may be in the form of hard-shelled capsules, which are normallyused for dry, powdered ingredients or miniature pellets, or soft-shelledcapsules, which are primarily used for oils and for active ingredientsthat are dissolved or suspended in oil. In certain embodiments, capsulesare made from aqueous solutions of gelling agents such as gelatin orplant polysaccharides. Other ingredients can be added to the gellingagent solution, such as plasticizers such as glycerin and/or sorbitol todecrease the capsule's hardness, coloring agents, preservatives,disintegrants, lubricants and surface treatment.

Dosage levels of a halogen compound and/or chalcogenide present in acomposition described herein may be varied as so to obtain an amount ofthe halogen compound and/or chalcogenide that is effective to achievedthe desired therapeutic effect for a particular subject, halogencompound, chalcogenide, and mode of administration, without being toxicto the subject.

In certain embodiments, a unit dosage form of a composition of theinvention formulated for oral delivery, e.g., a pill, tablet, caplet, orcapsule, comprises or consists of about 0.005 mg to about 5000 mg, about0.05 to about 1000 mg, about 0.5 to about 500 mg, about 0.5 to about 250mg, about 0.5 mg to about 100 mg, about 1 mg to about 500 mg, about 1 toabout 250 mg, about 1 mg to about 125 mg, about 1 mg to about 120 mg,about 1 to about 100 mg, about 2.5 mg to about 250 mg, about 2.5 toabout 100 mg, about 0.5 mg to about 50 mg, about 1 mg to about 50 mg,about 2.5 mg to about 50 mg, about 5 mg to about 50 mg, about 10 mg toabout 50 mg, about 1 to about 15 mg, about 1 to about 10 mg, or about 1mg, about 2 mg, about 5 mg, about 10 mg, or about 15 mg of a halogencompound, such as an iodide or NaI. The unit dosage form may furthercomprise a pharmaceutically acceptable carrier, excipient or diluent.

In particular embodiments, a unit dosage form of a composition of theinvention formulated for oral delivery, e.g., a pill, tablet, caplet, orcapsule, comprises or consists of less than 150 mg, less than 140 mg,less than 130 mg, less than 120 mg, less than 110 mg, less than 100 mg,less than 90 mg, less than 80 mg, less than 70 mg, less than 60 mg, lessthan 50 mg, less than 40 mg, less than 30 mg, less than 20 mg, less than10 mg, or less than 5 mg of a halogen compound, such as an iodide orNaI. The unit dosage form may further comprise a pharmaceuticallyacceptable carrier, excipient or diluent.

In certain embodiments, a unit dosage form may comprise a chalcogenide,alone or in combination with another active agent, such as a halogencompound. In specifically embodiments, the chalcogenide, e.g., sulfideor selenide, is present in the unit dosage form in the amount of about0.005 mg to about 5000 mg, about 0.05 mg to about 5000 mg, about 0.1 mgto about 5000 mg, about 0.005 mg to about 2000 mg, about 0.05 mg toabout 2000 mg, about 0.1 mg to about 2000 mg, about 0.5 mg to about 2000mg, about 1 mg to about 1000 mg, about 2 mg to about 500 mg, about 5 mgto about 500 mg, about 10 mg to about 100 mg, about 10 mg, about 20 mg,about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 100 mg, orabout 150 mg.

In particular embodiments of unit dosage forms suitable for oraladministration, the unit dosage form comprises both a halogen compoundand a chalcogenide. In certain embodiments, both the halogen compoundand the chalcogenide, e.g., sulfide or selenide, are in a reduced form.In one embodiment, the unit dosage form comprises a solid outer layercomprising the halogen compound, e.g., iodine, surrounding orencapsulating an inner region comprising a liquid solution comprisingthe chalcogenide. In particular embodiments, the unit dosage formcomprises a solid outer layer encapsulating an inner liquid regioncomprising the halogen compound and the chalcogenide. In certainembodiments, the encapsulated chalcogenide is protected from oxidationby the solid outer layer. In certain embodiments, the active agents arecomingled at the molecular level.

In particular embodiments of unit dosage forms suitable for oraladministration, the unit dosage form comprises both glutathione and achalcogenide. In certain embodiments, the chalcogenide, e.g., sulfide orselenide, is in a reduced form. In certain embodiments, the unit dosageform further comprises a halogen compound. In certain embodiments, theagents are comingled at the molecular level.

In particular embodiments of unit dosage forms suitable for oraladministration, the unit dosage form comprises both glutathione and ahalogen compound. In certain embodiments, the halogen compound, e.g.,iodine, is in a reduced form, e.g., iodide or iodate. In certainembodiments, the unit dosage form further comprises a chalcogenidecompound. In certain embodiments, the agents are comingled at themolecular level.

In various embodiments, compositions and unit dosage forms of theinvention may be formulated in any different manner suitable for adesired delivery route. Typically, formulations include allphysiologically acceptable compositions. Such formulations may include ahalogen compound, alone or in combination with another active agent,such as a chalcogenide, in combination with any physiologicallyacceptable carrier, diluent or excipient. Such formulations may includea chalcogenide, alone or in combination with another active agent, suchas a halogen compound, in combination with any physiologicallyacceptable carrier, diluent or excipient. Halogen compounds and/orchalcogenides may be formulated for administration with any biologicallyacceptable medium, including but not limited to water, buffered saline,polyol, or mixtures thereof. “Biologically acceptable medium” includesany and all solvents, dispersion media, and the like that may beappropriate for the desired route of administration of thepharmaceutical composition. Suitable biologically acceptable media andtheir formulations are described, for example, in the most recentRemington's Pharmaceutical Sciences (Remington's PharmaceuticalSciences. Mack Publishing Company, Easton, Pa., USA 1985).

Formulations, and unit dosages forms thereof, may contain suitablephysiologically acceptable carriers comprising excipients and/orauxiliaries that facilitate processing of the halogen compound and/orother active agent into preparations that can be used pharmaceutically.Formulations, and unit dosage forms thereof, may also include agentsthat increase or otherwise affect the bioavailability of the halogencompound and/or other active agent. As used herein, “bioavailability”refers to the effect, availability and persistence of the activeagent(s) after being administered to a subject.

Pharmaceutically acceptable carriers can be any pharmaceuticallyacceptable material, composition, or vehicle, including but not limitedto a liquid or solid filler, diluent, excipient, solvent orencapsulating material, involved in carrying or transporting the subjectagonists to an organ, or portion of the body. Each carrier must becompatible with the other ingredients of the formulation and notinjurious to the subject. Some examples of materials which can serve aspharmaceutically acceptable carriers include but are not limited tosugars, such as lactose, glucose and sucrose; starches, such as cornstarch and potato starch; cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate;tragacanth; malt; gelatin; talc; cocoa butter, waxes, animal andvegetable fats, paraffins, silicones, bentonites, silicic acid, zincoxide; oils, such as peanut oil, cottonseed oil, safflower oil, sesameoil, olive oil, corn oil and soybean oil; glycols, such as propyleneglycol; polyols, such as glycerin, sorbitol, mannitol and polyethyleneglycol; esters, such as ethyl oleate and ethyl laurate; agar; bufferingagents, such as magnesium hydroxide and aluminum hydroxide; alginicacid; pyrogen-free water; isotonic saline; Ringer's solution; ethylalcohol; phosphate buffer solutions; and any other compatible substancesemployed in pharmaceutical formulations.

Formulations can also include wetting agents; emulsifiers and lubricantssuch as sodium lauryl sulfate and magnesium stearate; coloring agents;release agents; coating agents; sweetening, flavoring, and/or perfumingagents; preservatives; and antioxidants. Formulations may alsoincorporate buffering agents and/or salts to aid absorption or stabilizethe halogen compound and/or chalcogenide. Other additives, such aschelating agents, enzymatic inhibitors, and the like, which wouldfacilitate the biological activity of the pharmaceutical composition mayalso be incorporated in the formulation.

The formulations may be presented in a unit dosage form and may beprepared by any methods known in the art. The amount of halogen compoundthat can be combined with a carrier to produce a single dosage form willgenerally be that amount of the halogen compound that produces atherapeutic effect.

Formulations suitable for oral administration may be in the form of asolid (capsules, cachets, pills, tablets, lozenges, powders, dragees,granules); or as a solution or a suspension in an aqueous or non-aqueousliquid; or as an oil-in-water or water-in-oil liquid emulsion; or as anelixir or syrup; or as pastilles (using an inert base, such as gelatinand glycerin, or sucrose and acacia); and/or as mouth rinses or washesand the like; or as a bolus, electuary or paste.

Solid formulations may have pharmaceutically acceptable carriers andextenders including but not limited to sodium citrate or dicalciumphosphate; starches; lactose; sucrose; glucose; mannitol; and/or silicicacid. Solid formulations can include additional components including butnot limited to binders such as carboxymethylcellulose, alginates,gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; humectants suchas glycerol; disintegrating agents such as agar-agar, calcium carbonate,potato or tapioca starch, alginic acid, certain silicates, and sodiumcarbonate; solution retarding agents such as paraffin; absorptionaccelerators such as quaternary ammonium compounds; wetting agents suchas cetyl alcohol and glycerol monostearate; absorbents such as kaolinand bentonite clay; lubricants such a talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, andmixtures thereof; and coloring agents. The formulation may also includebuffering agents, particularly when the halogen compound and/orchalcogenide is in the form of a capsule, tablet or pill.

Solid formulations may also include fillers for soft and hard-filledgelatin capsules using excipients such as lactose or milk sugars, aswell as high molecular weight polyethylene glycols and the like.

Solid formulations of compositions described herein, such as dragees,capsules, pills and granules, may optionally be scored or prepared withcoatings and shells, such as enteric coatings and other coatings. Soliddosage forms may also be formulated so as to provide slow or controlledrelease of the agent agent(s). Thus, solid formulations could includeany material that could provide a desired release profile of the activeagent(s), including but not limited to hydroxypropylmethyl cellulose invarying proportions, or other polymer matrices, liposomes and/ormicrospheres. Formulations of may also be formulated by including anembedding agent. Examples of embedding agents which can be used includebut are not limited to polymeric substances and waxes. The halogencompound and/or chalcogenide may also be in microencapsulated form, ifappropriate, with one or more of the above-described excipients.

Liquid dosage formulations for oral administration of the halogencompound and/or chalcogenide may include pharmaceutically acceptableemulsions, microemulsions, solutions, suspensions, syrups and elixirs.In addition to the halogen compound and/or chalcogenide, the liquiddosage formulations may contain inert diluents commonly used in the art,including but not limited to water or other solvents; solubilizingagents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol; oils (in particular, cottonseed, groundnut,corn, germ, olive, castor and sesame oils); glycerol; tetrahydrofurylalcohol; polyethylene glycols; and fatty acid esters of sorbitan, andmixtures thereof.

The present invention further includes stable liquid pharmaceuticalcompositions formulated for parenteral administration, e.g., intravenousadministration or administration by infusion. In certain embodiments,the stable liquid pharmaceutical compositions comprise a reduced form ofa chalcogenide and/or a reduced form of a halogen compound. Inparticular embodiments, the composition further comprises glutathione.In other embodiments, a stable liquid pharmaceutical compositionsformulated for parenteral administration comprises a goitrogen or acompound that inhibits or impedes thyroid hormone synthesis, activity oruptake. In particular embodiments, a stable liquid pharmaceuticalcompositions formulated for parenteral administration comprises achalcogenide, e.g., selenide, and glutathione. In particularembodiments, a stable liquid pharmaceutical compositions formulated forparenteral administration comprises a halogen compound, e.g., iodide,and glutathione. In particular embodiments, a stable liquidpharmaceutical compositions formulated for parenteral administrationcomprises selenide, iodide, and glutathione. The selenide, iodide andglutathione may be present at a concentration described herein, or inamount sufficient or appropriate to deliver an amount described hereinto a subject. The concentration of each active agent in the compositionmay be readily determined based on the desired amount of each activeagent to be delivered to a subject in need thereof.

In particular embodiments, a composition of the invention comprises achalcogenide, a halogen compound, and glutathione. In certainembodiments, the chalcogenide is sulfide or selenide (or a reduced formthereof), and the halogen compound is iodide or iodate. In particularembodiments, the chalcogenide is selenide. In particular embodiments,the halogen compound is iodide. Thus, in one embodiment, a compositionof the present invention comprises selenide, iodide and glutathione. Thecomposition may be formulation for intravenous administration oradministration by infusion. The composition may be formulated as astable composition for intravenous administration or administration byinfusion. In particular embodiments, the composition comprises selenideat a concentration of about 1 mM to about 1 M or about 10 mM to about500 mM, iodide at a concentration of about 1 mM to about 1M or about 10mM to about 500 mM, and glutathione at a concentration of about 1 mM toabout 500 mM or about 10 mM to about 500 mM. In particular embodiments,the composition is contained within an oxygen-impermeable container, andmay be under nitrogen or argon gas. In particular embodiments, theamount of composition present in the container is a unit dosage amountcomprising or consisting of a suitable dosage amount for administrationto a subject in need thereof.

Formulations of halogen compound and/or chalcogenides for parenteraladministration may comprise a halogen compound and/or a chalcogenide incombination with one or more pharmaceutically acceptable isotonicaqueous or nonaqueous solutions, dispersions, suspensions or emulsions,or powders which may be reconstituted into sterile injectable solutionsor dispersions just prior to use. Parenteral formulations may containantioxidants; buffers or solutes which render the formulation isotonicwith the blood of the intended subject; bacteriostats; suspending; orthickening agents.

Injectable depot formulations comprising a halogen compound and/or achalcogenide can be made by forming microencapsulated matrices of thehalogen compound and/or chalcogenides in biodegradable polymers.Examples of biodegradable polymers include, but are not limited topolylactide-polyglycolide, poly(orthoesters) and poly(anhydrides). Theratio of active agent to polymer and the nature of the particularpolymer employed can affect the rate of active agent release. Depotinjectable formulations can also be prepared by entrapping the drug inliposomes or microemulsions.

The present invention further includes kits comprising compositions orunit dosage forms of the present invention. In certain embodiments, suchkits comprise one or more containers to store the compositions of thepresent invention. In one embodiment, the composition is stored in thecontainer under an inert or noble gas, and the container is a sealed andhas an oxygen impermeable light-protective container (e.g., an ambervial).

In particular embodiments, a composition of the present inventioncomprises a carrier. In certain embodiments, the halogen compound isassociated with the carrier, and in other embodiments, both the halogencompound and one or more additional active agents present in thecomposition are also associated with the carrier. The halogen compoundand one or more additional active agents may be associated covalently ornon-covalently. In certain embodiments, the carrier is a biocompatiblecarrier, which may be, e.g., biodegradable or inert. In certainembodiments, the carrier is a polypeptide, such as, e.g., an albumin,e.g., human albumin. Examples of other carriers that may be usedaccording to the present invention include, but are not limited toplasma, serum, alpha-2-macroglobulin, and immunoglobulin.

Methods of Preparing Compositions

Compositions comprising halogen compounds, including reduced forms ofhalogen compounds, such as iodide and bromide, may be prepared by anymeans known and available. In certain embodiments, a halogen compound isdissolved in water or a suitable buffer, such as a NaCl buffer.

Certain chalcogenide compounds (e.g., hydrogen sulfide, hydrogenselenide), are not stable in the presence of oxygen due to their abilityto react chemically with oxygen, leading to their oxidation and chemicaltransformation. Accordingly, oxygen may be removed from solutions usedin these methods, using methods known in the art, including, but notlimited to, application of negative pressure (vacuum degassing) to theor solution, or contacting the solution with a reagent which causesoxygen to be bound or “chelated”, effectively removing it from solution.In general, methods of preparing compositions used according to thepresent invention may include limiting oxygen content in each aspect ofmanufacturing and storage.

WO2008/043081 describes methods of preparing stable, liquid compositionscomprising chalcogenides or salts thereof, including those comprisingreduced forms of chalcogenides.

U.S. Provisional Application No. 61/159,311 also describes methods ofpreparing a composition comprising a reduced form of a chalcogenide,e.g., a chalcogenide comprising selenium or sulfur is the −2 valencestate, e.g., selenide or sulfide.

In particular embodiments of any of the methods of producing a stablecomposition described herein, the reduced form of the chalcogenide is inthe minus 2 oxidation or valence state. In certain embodiments, thereduced form of the chalcogenide is H₂Se, Na₂Se, NaHSe, or HSe— anion.

In one embodiment, a composition comprising a reduced form of achalcogenide is produced as follows: admixing a chalcogen, or an acid ofa chalcogen, and a reducing agent in a reduced oxygen environment underconditions and for a time period sufficient to allow oxidation of amajority of the reducing agent and reduction of a majority of thechalcogen. In certain embodiments, the conditions include a temperatureof about room temperature. In particular embodiments, the time period isabout one hour, about two hours, about three hours, or overnight, e.g.,about 12 hours. In particular embodiments, the time period is finishedor ends when the admixture solution appears visibly clear or bubbling(due to hydrogen generation from the borohydride reacting with water)ceases or is no longer observed.

In particular embodiments, the chalcogen is sulfur or selenium, and thereduced form of a chalcogenide comprises sulfur or selenium in the −2oxidation state. In certain embodiments, the acid of a chalcogen isselenous acid or sodium selenite or elemental selenium, and the reducedform of a chalcogenide comprises sulfur or selenium in the −2 oxidationstate.

In particular embodiments, the reducing agent has a reductionpotential)(E° less than or equal to about 0.4 V. In one embodiment, thereducing agent is sodium borohydride (NaBH₄). In certain embodiments,the molar ratio of the reducing agent to the chalcogen, or the acid of achalcogen, is about 5:1 to about 0.5:1, or about 3:1 to about 1:1. Inone embodiment, the molar ratio of the reducing agent to the chalcogen,or the acid of a chalcogen, is about 2:1.

In particular embodiments, the reducing agent is sodium borohydride,said chalcogen is sulfur or selenium, and said acid of a chalcogen isselenious acid. In one particular embodiment, the reducing agent issodium borohydride, the chalcogen is selenium, and the molar ratio ofsodium borohydride to selenium is about 2:1. In particular embodiments,the selenium is present in an amount of about 1 mM to about 10 M, orabout 1 mM to 1M (79 mg/L to 79 g/L). In particular embodiments, thesodium borohydride is present in a 1M solution in water.

In certain embodiment, the method is performed in a reduced oxygen oroxygen-free environment. In certain embodiments, the method is performedunder an inert or noble gas. In certain embodiments, the method isperformed under nitrogen. For example, the nitrogen may be perfused intosaid reduced oxygen environment. In certain embodiments, the nitrogen isperfused at a rate of about 100 cc/min. In particular embodiments, thereduced oxygen environment is a container having an oxygen-freeenvironment. In particular embodiments, the container is a syringe, i.v.bag, tube or a vial. In certain embodiments, the container comprises aclosable port of entry or a resealable port. In one embodiment, thecontainer is a sealable tube comprising a rubber septum, e.g., a Hungatetube.

In particular embodiments of this method, the perfusion of nitrogen intoand out of the container, e.g., tube, occurs via two needles that passthrough the septum of the container, wherein one of the two needles isused as the port to bring nitrogen into the tube, and wherein the secondof the two needles is used to take nitrogen out of the tube.

In particular embodiments, the method further includes heating theadmixture after said time period. In particular embodiments, the heatingis continued until any observed bubbling ceases. The methods may alsoinclude cooling the admixture after the heating. Cooling may beperformed, e.g., by placing the admixture on ice. In certainembodiments, cooling is continued until sodium borate precipitates fromthe admixture solution.

These methods may also further comprise centrifuging the admixturesolution to separate a supernatant from the precipitated sodium borate,and removing said supernatant, wherein the supernatant comprises saidstable composition comprising said reduced chalcogenide.

In particular embodiments, the methods comprise acidifying the admixturewith an acid, wherein said acid is reducing and not volatile, andbubbling hydrogen selenide gas or hydrogen sulfide gas through asolution, wherein the solution has a pH greater than 3.9. Withoutwishing to be bound to any particular theory, it is believed that if anon volatile acid (e.g. phosphorus acid) is added to a solution ofreduced chalcogenide that may have unwanted non volatile compounds inthe solution, it will lower the pH to a value less than the pK for thechalcogenide, thus producing a gas form (e.g. H₂S or H₂Se). The gas formis able to be blown out of the solution by passing nitrogen through themixture and carried to a second solution that has a pH above the pKrendering the chalcogenide into an ionic form that is no longer a gas(i.e. trapping the chalcogenide).

In certain embodiment, the acid is phosphorous acid and said solution isphosphate buffered saline (PBS). This permits the hydrogen selenide orhydrogen sulfide gas to be trapped in a vessel.

In another embodiments, the present invention includes another method ofproducing a composition comprising a reduced form of a chalcogenide, themethod comprising admixing elemental selenium (Se) or sulfur (S) andsodium hydride in a solution comprising mineral oil or tetrahydrofuran(THF), thereby producing a stable composition comprising sodiumhydroselenide or sodium sulfide. In certain embodiments, the method alsocomprises adding water to the admixture solution, thereby removingsodium hydride. In particular embodiments wherein the solution comprisesmineral oil, the method further comprises removing the aqueous phase ofthe admixture solution, wherein the reduced chalcogenide is present inthe aqueous phase. In particular embodiments wherein the solutioncomprises THF, the method further comprises removing said THF by boilingsaid admixture solution at about 70° C.

In various embodiments, methods of preparing compositions of the presentinvention further include adjusting the pH of the composition. Incertain embodiments, the pH is adjusted by the addition of one or moreof hydrogen chloride, carbon dioxide, nitrogen, or hydrogen sulfide. Inanother embodiment, the pH is adjusted by dissolving nitrogen, carbondioxide, hydrogen selenide, or hydrogen sulfide into the composition orany combination thereof.

In certain embodiments, once produced, in various embodiments, acomposition is stored in an impermeable container, e.g., an oxygenimpermeable container. This is particularly desirable to preventoxidation of the reduced form of halogen compound or chalcogenide.Impermeable containers are known to those skilled in the art andinclude, but are not limited to, “i.v. bags” comprising a gasimpermeable construction material, or a sealed glass vial. In particularembodiments, the impermeable container comprises an oxygen impermeablematerial having an oxygen transmission coefficient less than 10⁻¹⁰[cm³(STP)/cm/(cm²+s+Pa)], wherein STP=standard temperature and pressure(25 degrees centigrade and pressure 1 atmosphere); PA=pascals, ands=second. For example, the walls of the container may comprise a layerof an oxygen impermeable polymer. Exemplary oxygen impermeable polymersinclude but are not limited to: silicon rubber, natural rubber, lowdensity poly ethylene (LDPE), polystyrene (PS), polyethylene (PE),polycarbonate (PC), polyvinyl acetate (PVAc), amorphous polyethyleneterephthalate (APET), polyvinly chloride (PVC), nylon 6 (Ny6), polyvinylfluoride (PVF), polyvinylidene chloride (PVdC), polyacetonitrile (PAN),ethylene vinyl alcohol (EVOH), and polyvinyl alcohol (PVA). In certainembodiments, the oxygen transmission coefficient of said polymer is lessthan 10⁻¹⁰ [cm³(STP)/cm/(cm²+s+Pa)]. In particular embodiments, thewalls of the container comprise multiple layers of one or more oxygenimpermeable polymers.

In additional embodiments, the container comprises one or moreresealable or closable ports of entry. In certain embodiments, thecontainer comprises two or more resealable or closable ports of entry.As noted, in specific embodiments, the container is a bottle, a bag, atube, a vial, or a syringe. In certain embodiments, the container is anintravenous bag or a syringe. In particular embodiments, the containeris a sealable tube comprising a rubber septum, e.g., a Hungate tube.

In related embodiments, the device further comprises a delivery meanscoupled to the container through a resealable or closable port of entry.In particular embodiments, the delivery means is configured tointravenously deliver a solution from the container to a subject in needthereof. For example, the delivery means may be a needle or a cannula.In certain embodiments, the delivery means comprises a reduced oxygen oroxygen-free environment or is present under a reduced oxygen oroxygen-free environment.

In particular embodiments of storage or delivery devices of the presentinvention, the compound is a halogen compound or a chalcogenide. Incertain embodiments, the compound is a reduced form of a halogencompound or a chalcogenide, e.g., an iodide, a bromide, NaI, KI, HI,CaI, AGI, H₂Se, Na₂Se, NaHSe, or HSe— anion or others described herein.In certain embodiments, the device comprises a composition of thepresent invention.

To prevent exposure to air in the gas-tight storage container, an inertor noble gas, such as nitrogen or argon, may be introduced into acontainer containing a composition of the present invention prior toclosure.

In other related embodiments, compositions are stored in alight-resistant or a light-protective container or vial, such as anamber vial. The composition may be packaged in a glass vial. It may befilled to a slight over-pressure in an inert atmosphere, e.g., nitrogen,to prevent/slow oxidative breakdown of the composition, and may becontained in a form such that ingress of light is prevented, therebypreventing photochemical degradation of the composition. This may beachieved using an amber vial. Additional container systems that permit asolution to be stored in an oxygen-free environment are known, as manyintravenous solutions are sensitive to oxygen. For example, a glasscontainer that is purged of oxygen during the filling and sealingprocess may be used. In another embodiment, flexible plastic containersare available that may be enclosed in an overwrap to seal againstoxygen. Basically, any container that prevents oxygen from interactingwith the stable composition may be used (see, e.g., U.S. Pat. No.6,458,758). In one embodiment, the container includes one or more oxygenscavenger. For example, the oxygen scavenging composition can be appliedas a coating or lining upon the inside surface of the product supportingor retaining means to function as a barrier to oxygen permeation (see,e.g., U.S. Pat. No. 5,492,742).

In particular embodiments, a container or vial may comprise a unitdosage of a composition of the present invention. In certainembodiments, the unit dosage form comprises or consists of an effectiveamount of the composition to treat or prevent a disease, condition, orinjury, including any of those described herein, in a subject.

In particular embodiments, the present invention includes a container,such as a saline bag, that includes a premixed liquid composition of ahalogen compound, e.g., an iodide or bromide, wherein the amount ofpremixed liquid composition constitutes a dosage useful in treating orpreventing a disease, condition, or injury, including any of thosedescribed herein, in a subject in need thereof, and one or morepharmaceutically acceptable carriers, diluents, or excipient. Inparticular embodiments, the liquid composition is sterile.

In particular embodiments, the present invention includes a container,such as a vial, that includes a dry composition of a halogen compound,e.g., an iodide or bromide, wherein the amount of dry compositionconstitutes a dosage useful in treating or preventing a disease,condition, or injury, including any of those described herein, in asubject in need thereof. The dry composition may be reconstituted, e.g.,with a pharmaceutically acceptable carrier, diluent, or excipient, e.g.,sterile water, prior to delivery to a subject in need thereof.

In certain embodiments, compositions comprising a halogen compound andan active agent used for the treatment of chronic heart failure may beprepared by admixing a reduced form of a halogen compound, e.g., iodide,and the active agent used for the treatment of chronic heart failure. Incertain embodiments, one or more pharmaceutically acceptable excipients,diluents or carriers are also admixed.

Methods of Using Halogen Compounds and Other Compositions

In certain embodiments, a composition of the present invention is usedto treat or prevent an injury or a disease in a biological material,e.g., a subject, e.g., a mammal, such as a human. In particularembodiments, a composition of the present invention is used to treat asubject (or biological material) prior to, during, or after exposure toan ischemic or hypoxic condition, or reperfusion. Biological materialmay be treated in vivo, ex vivo, or in vitro. In specific embodiments, acomposition of the invention is used to treat or prevent injury causedby a heart attack or infarct resulting from a heart attack or stroke. Inspecific embodiments, a composition of the invention is used to treat orprevent injury caused by a cell, tissue, or organ transplant. A varietyof diseases, injuries, and conditions are described herein, all of whichmay be treated by a composition of the present invention, e.g., acomposition comprising a halogen compound such as iodide, or acomposition comprising a chalcogenide such as sulfide or selenide. Inparticular embodiments, the composition is a stable formulationformulated to maintain the halogen compound or chalcogenide in itsreduced state.

In certain embodiments, the present invention includes methods oftreating or preventing a disease or injury in a biological material inneed thereof, comprising providing to the biological material aneffective amount of a composition of the present invention. Inparticular embodiments, the composition comprises an effective amount ofone or more halides and/or one or more chalcogenides, and optionally oneor more additional active agents, including any of those describedherein. In particular embodiments, the halide or halogen compoundcomprises iodine, e.g., iodide or iodate. In particular embodiments, thechalcogenide comprises sulfur or selenium, e.g., sulfide or selenide. Inparticular embodiments, the composition also comprises glutathione oranother reducing agent.

In one embodiment, compositions of the present invention are used totreat subjects who have undergone, are undergoing, or who aresusceptible to a disease, injury, trauma or critical care treatment.Thus, the present invention includes a method of treating or preventinga disease, injury, or trauma, including an injury related to criticalcare treatment, in a subject in need thereof, comprising providing tothe subject an effective amount of a composition of the presentinvention. In particular embodiments, the composition comprises aneffective amount of one or more halides and/or one or morechalcogenides, and optionally one or more additional active agents,including any of those described herein. In particular embodiments, theinjury may be caused by external insults, such as burns, wounds,amputations, gunshot wounds, or surgical trauma, abdominal surgery,prostate surgery; internal insults, such as septic shock, stroke orcardiac arrest, heart attack, chronic heart failure, e.g., that resultin the acute reduction in circulation; or reductions in circulation dueto non-invasive stress, such as exposure to cold or radiation. On acellular level, injury often results in exposure of cells, tissuesand/or organs to hypoxia, thereby resulting in induction of programmedcell death, or “apoptosis.” In particular embodiments, injury resultsfrom reperfusion of oxygen into cells, a tissue, an organ, or a mammalfollowing a hypoxic or ischemic event. In particular embodiments,compositions and methods of the present invention, including thoserelated to either or both halide and/or chalcogenides, are used to treator prevent surgical adhesion, e.g., abdominal, pelvic, and heartsurgical adhesions. In certain embodiments, an adhesion is a band ofscar tissue that binds two parts of tissue or organs together. Theadhesion may develop when the body's repair mechanisms respond to tissuedisturbance, e.g., surgery, infection, trauma or radiation. Althoughadhesions can occur anywhere, common locations are within the abdomen,pelvis and heart. In certain embodiments, compositions and methods ofthe present invention, including those related to either or both halideand/or chalcogenides, are used to treat or prevent injury associatedwith stem cell engraftment, and/or to promote stem cell engraftment orreduce stem cell rejection, e.g., of allogeneic or autologous stem cellsintroduced to a subject.

In one embodiment, the present invention contemplates contacting cells,tissues, organs, limbs, and even whole organisms, with an effectiveamount of a composition of the present invention as a way of protectingthem from a detrimental effect of injury or disease. The presentinvention also contemplates methods for inducing tissue regeneration andwound healing by inhibition/prevention/delay of biological processesthat may result in delayed wound healing and tissue regeneration. Incertain embodiments, the present invention includes methods ofprotecting a biological material, or preventing or inhibiting injury toa biological material in need thereof, comprising providing to thebiological material an effective amount of a composition of the presentinvention. In particular embodiments, the composition comprises aneffective amount of one or more halides and/or one or morechalcogenides, and optionally one or more additional active agents,including any of those described herein. In this context, in scenariosin which there is a substantial wound to the limb or organism,contacting the biological matter with a composition of the presentinvention aids in the wound healing and tissue regeneration process bymanaging the biological processes that inhibit healing and regeneration.In addition to wound healing, methods of the invention can beimplemented to prevent or treat trauma such as cardiac arrest or stroke,or hemorrhagic shock. The invention has importance, e.g., with respectto the risk of trauma from emergency surgical procedures, such asthoroacotomy, laparotomy, and splenic transaction or cardiac surgery,aneurysm, surgery, brain surgery and the like.

In certain embodiments, methods of the present invention can beimplemented to enhance survivability or inhibit/prevent ischemic injuryor inhibit/prevent reperfusion injury, e.g., resulting from cardiacarrest or stroke. Accordingly, in one embodiment, the present inventionincludes methods of enhancing survivability or reducing ischemic orreperfusion injury by providing to a biological material diagnosed withor at risk of ischemic injury or inhibit/prevent reperfusion injury,e.g., resulting from cardiac arrest or stroke, with a composition of theinvention, e.g., a composition comprising a halogen compound, optionallyin combination with one or more additional active agents, such as achalcogenide. This may be, e.g., in a subject suffering from or at riskof cardiac arrest or stroke, comprising providing an effective amount ofa composition of the invention to the subject before, after, or bothbefore and after myocardial infarction, cardiac arrest or stroke.

“Ischemia” refers to a restriction in blood supply to tissues, causing ashortage of oxygen and glucose needed for cellular metabolism. It can becaused by problems with blood vessels, leading to damage to the affectedtissue. In certain embodiments, it results from vasoconstriction,thrombosis, myocardial infarct, stroke, or embolism. As used herein,“reperfusion injury” refers to tissue damage resulting from bloodreturning to tissue after a period of ischemia or lack of oxygen. It isbelieved that the absence of oxygen and nutrients from blood during theischemic period creates a condition in which the restoration ofcirculation results in inflammation and oxidative damage through theinduction of oxidative stress. In certain embodiments, the methodresults in a reduced infarct size as compared to the size in the absenceof treatment with the stable composition.

In certain embodiments, methods of the present invention includepre-treating a biological material, e.g., a subject, with a compositionof the present invention prior to an ischemic, hypoxic, or reperfusioninjury or event or disease insult. These methods can be used when aninjury or disease with the potential to cause ischemia or hypoxia isscheduled or elected in advance, or predicted in advance to likelyoccur. Examples include, but are not limited to, major surgery whereblood loss may occur spontaneously or as a result of a procedure,cardiopulmonary bypass in which oxygenation of the blood may becompromised or in which vascular delivery of blood may be reduced (as inthe setting of coronary artery bypass graft (CABG) surgery), or in thetreatment of cell, tissue or organ donors (or donor cells, tissues, ororgans) prior to removal of donor cells, tissues, or organs fortransport and transplantation into a recipient in need of a cell,tissue, or organ transplant. In various embodiments, the donor cell,tissue, or organ is autologous or heterologous to the graft ortransplant recipient. Examples include, but are not limited to, medicalconditions in which a risk of injury or disease progression is inherent(e.g., in the context of unstable angina, following angioplasty,bleeding aneurysms, hemorrhagic strokes, following major trauma or bloodloss), or in which the risk can be diagnosed using a medical diagnostictest.

In certain embodiments, compounds, compositions and methods of thepresent invention are used to treat, inhibit, reduce or prevent any ofthe following diseases or conditions, or an injury related to orresulting from any of the following treatments: transplantation (e.g.,kidney transplant, cadaveric kidney transplant, liver transplant, boweltransplant, lung transplant, or flap transplant); ileus (e.g., failureof peristalsis and post-operative ileus); graft vs. host disease;radiation-induced lung injury; ventilator induced lung injury; cataract(e.g., incipient cataract, senile cataract, or incipient senilecataract); Hanta pulmonary virus (HPV); Ebola virus; Salmonellainfection, bacterial infection; ischemic birth (e.g., Hypoxic IschemicEncephalopathy (HIE) or birth asphyxia); peripheral vascular disease;permanent ischemia; ST segment elevation myocardial infarction (STEMI);hearing loss (e.g., loud noise hearing loss or acute hearing loss);status epilepticus (epilepsy), superior mesenteric arteryischemia/reperfusion injury (e.g., ischemia-reperfusion injury followingsuperior mesenteric artery occlusion); sickle cell anemia or sickle cellcrisis; or contrast-induced nephropathy. In certain embodiments, any ofthese diseases, disorder or injuries are treated with a compositioncomprising a chalcogenide, or a composition comprising a halide. Inparticular embodiments, the composition comprising a chalcogenidecomprises sulfide or selenide, e.g., in a reduced form, including any ofthe compositions described herein, such s those that maintain thesulfide or selenide in a reduced form.

In certain embodiments, the invention comprises treating or preventingradiocontrast agent induced kidney injury in a subject in need,comprising providing to the subject an effective amount of a compositioncomprising a halogen (e.g., iodide) or a chalcogenide (e.g., sulfide orselenide). Radiocontrast agent induced kidney injury (contrast-inducednephropathy) represents a significant clinical problem and has beenassociated with increased patient mortality and longer hospitalizations.Radiocontrast agent administration, conducted in conjunction withimaging of the coronary arteries, the peripheral arteries or the kidneysis a common medical intervention. However, radiocontrast agentadministration often leads to significant kidney damage in a subset ofpatients with certain risk factors. For example, patients who areelderly, have an underlying disease (e.g., diabetes or atherosclerosis)and/or have a borderline impairment in their renal function areespecially predisposed to kidney injury. In one embodiment, theradiocontrast agent is administered before or during kidney diagnosticimaging. In another exemplary embodiment, the radiocontrast agent isadministered before or during heart blood vessel diagnostic imaging. Inanother exemplary embodiment of the method, the radiocontrast agent isan iodinated radiocontrast agent. In another exemplary embodiment of themethod, the subject (e.g., human patient in need of treatment thereof)has a pre-existing impairment of renal function prior to administrationof the radiocontrast agent. In another exemplary embodiment of themethod, the human has a baseline creatinine level greater than or equalto 120 μmol/L, a glomerular filtration rate of less than 60 mL/min per1.73 m² and/or a creatinine clearance of less than 60 mL/min. In anotherexemplary embodiment of the method, the human has one of more conditionsselected from type 1 or 2 diabetes mellitus, atherosclerosis, congestiveheart failure, an intraarterial balloon pump, anemia, a systolic bloodpressure of less than 80 mm Hg, an age greater than 50 years, aglomerular filtration rate of less than 60 mL/min per 1.73 m² andreduced intravascular volume. In another exemplary embodiment of themethod, the radiocontrast agent induced kidney injury is radiocontrastagent induced nephropathy or acute renal dysfunction.

Thus, in certain embodiments, the present invention includes a method oftreating, preventing, reducing, inhibiting (or reducing or inhibiting aninjury associated with) any of the disorders or procedures describedherein in a biological material (e.g., a subject), comprising providingan effective amount of a compound or composition described herein to thebiological material (e.g., the subject).

In certain embodiments, the present invention includes a method oftreating, preventing, reducing, inhibiting chronic heart failure in asubject in need thereof, comprising providing an effective amount of acompound or composition described herein to the subject (e.g., thesubject). In particular embodiments, the subject has been diagnosed ashaving or being susceptible to chronic heart failure, e.g., systolicheart failure. CHF may be diagnosed based on the history of the symptomsand a physical examination with confirmation by echocardiography. Inaddition, blood tests, electrocardiography, and chest radiography may beuseful to determine the underlying cause. In particular embodiments, thesubject has been diagnosed with chronic heart failure due to leftventricular dysfunction or systolic heart failure. In particularembodiments, the subject has undergone a heart attack, e.g., within theone day, one week, one month, 6 months, one year, or 5 years prior tothe commencement of the treatment. In certain embodiments, the subjectis provided with a halogen, such as an iodine-containing halogen, e.g.,iodide. In particular embodiments, the subject is provided with apharmaceutical composition comprising a reduced form of a halogencompound, e.g., a reduced form of iodine, such as iodide or iodiate, anda pharmaceutically acceptable carrier, diluent, or excipient. In certainembodiments, the pharmaceutical composition further comprisesglutathione. In certain embodiments, the glutathione is present in anamount sufficient to maintain the halogen or halogen compound in itsreduced state. In particular embodiments, the subject is provided withany of the compositions, e.g., pharmaceutical compositions, or unitdosage forms described herein via any of the routes of administrationdescribed herein. In particular embodiments, the subject is providedwith the halogen compound orally or intravenously.

In particular embodiments of methods of treating or preventing CHF,e.g., systolic heart failure, a halogen compound is provided to thesubject in combination with one or more other active agents used totreat CHF. In certain embodiments, the subject is provided an effectiveamount of a first pharmaceutical composition comprising a reduced formof a halogen compound, e.g., a reduced form of iodine, such as an iodideor iodiate, and a pharmaceutically acceptable carrier, diluent, orexcipient, and an effective amount of a second pharmaceuticalcomposition comprising another active agent used to treat CHF. It isunderstood that the effective amount of each pharmaceutical compositionwhen used in combination may be less than the effective amount of one orboth compositions when they are used alone. In certain embodiments, thetwo pharmaceutical compositions act synergistically to treat or preventCHF.

In certain embodiments, the another active agent used to treat CHF,e.g., CHF due to left ventricular dysfunction, is an angiotensinconverting enzyme (ACE) inhibitors or a beta blocker. In otherembodiments, the another active agent is an aldosterone antagonist, anangiotension receptor blocker, or hydralazine with a nitrate. Inadditional embodiments, the another active agent is an anticoagulant, anantiplatelet therapy, an angiotensin II receptor blocker, a calciumchannel blocker, a diuretic, a vasodilator, or a statin. In particularembodiments, a biological material, e.g., a mammalian cell, tissue,organ or a mammal, is pre-treated with a composition of the invention,e.g., a composition comprising a halogen compound, prior to an ischemic,hypoxic or reperfusion event or injury. In particular embodiments, thebiological material is contacted with the composition for at least orabout one hour, at least or about two hours, at least or about fourhours, at least or about six hours, at least or about eight hours, atleast or about ten hours, at least or about 12 hours, at least or about16 hours, at least or about 20 hours, at least or about 24 hours, atleast or about 36 hours, at least or about 48 hours, at least or aboutthree days, at least or about four days, at least or about five days, atleast or about one week, at least or about two weeks, at least or aboutone month, between one hour and one week, between one hour and 48 hours,between one hour and 24 hours, between two hours and one week, betweentwo hours and 48 hours, between two hours and 24 hours, between fourhours and one week, between four hours and 48 hours, between four hoursand 24 hours, between twelve hours and one week, between twelve hoursand 48 hours, between twelve hours and 24 hours, between 24 hours andone week, or between 24 hours and 48 hours prior to or immediately priorto the ischemic, hypoxic or reperfusion event or injury. In certainembodiments, there is a period of time after treatment and immediatelyprior to the ischemic, hypoxic or reperfusion event or injury duringwhich the biological matter is not contacted with the composition of thepresent invention, e.g., a period of time of about five minutes, aboutten minutes, about 20 minutes, about 30 minutes, about one hour, abouttwo hours, about four hours, about eight hours, about 12 hours, or about24 hours. In particular embodiments, the biological material, e.g.,cell, tissue or organ, is grafted or transplanted to a recipient, eitherwhile being treated or after being treated with a composition of thepresent invention. In particular embodiments, the grafter ortransplanted biological material is autologous or heterogonous to therecipient.

Relatedly, additional embodiments of the invention concern enhancingsurvivability and preventing irreversible tissue damage from blood lossor other lack of oxygenation to cells or tissue, such as from lack of anadequate blood supply. In certain embodiments, the present inventionincludes methods of enhancing survivability or preventing tissue damagefrom blood loss or other lack of oxygenation to cells or tissue in abiological material in need thereof, comprising providing to thebiological material an effective amount of a composition of the presentinvention. In particular embodiments, the composition comprises aneffective amount of one or more halides and/or one or morechalcogenides, and optionally one or more additional active agents,including any of those described herein. Lack of adequate blood supplymay be the result of, for example, actual blood loss, or it may be fromconditions or diseases that cause blockage of blood flow to cells ortissue, that reduce blood pressure locally or overall in an organism,that reduce the amount of oxygen that is carried in the blood, or thatreduces the number of oxygen carrying cells in the blood. Conditions anddiseases that may be involved include, but are not limited to, bloodclots and embolisms, cysts, growths, tumors, anemia (including sicklecell anemia), hemophilia, other blood clotting diseases (e.g., vonWillebrand, or ITP), and atherosclerosis. Such conditions and diseasesalso include those that create essentially hypoxic or anoxic conditionsfor cells or tissue in an organism because of an injury, disease, orcondition.

In one embodiment, the present invention provides methods andcompositions to enhance the survivability of or reduce or prevent injuryor damage to biological material (e.g., a mammal or tissue or organwithin a mammal) undergoing hemorrhagic shock or undergoing reperfusion,which include contacting the biological material at risk of or in astate of hemorrhagic shock with an effective amount of a composition ofthe present invention as soon as practical, e.g., within one hour of theinjury. This method allows for the subject to be transported to acontrolled environment (e.g., surgery), where the initial cause of theinjury can be addressed, and then the patient can be brought back tonormal function in a controlled manner. For this indication, the firsthour after injury, referred to as the “golden hour,” is crucial to asuccessful outcome.

In various other embodiments, the methods and compositions of thepresent invention may be used in the treatment or prevention ofneurodegenerative diseases associated with ischemia, hypoxia orreperfusion, in the treatment of hypothermia, in the treatment ofhyperproliferative disorders, and in the treatment of immune disorders.In various other embodiments, the biological condition is any one orcombination of the following: neurological disease, cardiovasculardisease, metabolic disease, infectious disease, lung disease, geneticdisease, autoimmune disease, and immune-related disease. In certainembodiments, the methods and compositions of the present invention areused to enhance the survivability of ex vivo biological matter subjectedto hypoxic or ischemic conditions, including, e.g., isolated cells,tissues and organs. Specific examples of such ex vivo biologicalmaterial include platelets and other blood products, as well as cells,tissues and organs to be transplanted, e.g., for autologous orheterologous transplantation. In various embodiments, biologicalmaterial for transplantation is treated with a composition describedherein while still in the donor, after removal from the donor, or aftertransplantation into the recipient, or any combination thereof. Inparticular embodiments transplant recipient is treated with acomposition described herein before, during or after transplantation ofa biological material, e.g., cells, tissue or organ.

Methods of the present invention may be used to regulate the reducingenvironment of blood, a tissue, or cells, including, e.g., those presentat the site of injury or disease, such as, e.g., a myocardial infarct.In certain embodiments, the present invention includes methods ofregulating the reducing environment in a biological material, comprisingproviding to the biological material an effective amount of acomposition of the present invention. In particular embodiments, thecomposition comprises an effective amount of one or more halides and/orone or more chalcogenides, and optionally one or more additional activeagents, including any of those described herein. Without wishing to bebound to any particular theory, it is understood that under normalconditions, there exists in the red cells a constant ratio of reduced tooxidized glutathione of about 99:1. Under these conditions, anions suchas selenite and iodate enter red cells via the anion transporter/band 3protein on the plasma membrane, and upon being exposed to the highlyreduced glutathione become transformed/reduced to selenide and iodide.An additional by product of this reaction is the production of a smallamount of oxidized glutathione. This oxidized glutathione is re-reducedusing energy generated from the oxidation of glucose via the pentosephosphate pathway. The selenide and iodide leave via the aniontransporter/band 3 protein and have the effect of moving the reducingpower of the red cell to other parts of the body. When this pathwaybecomes overwhelmed, such as, e.g., during injury or disease, the ratioof reduced to oxidized glutathione falls, e.g., below 90:10, below80:10, below 70:10 below 60:10 or as low as 50:50 following a heartattack. In addition, it is thought that the amount of reduced anions,such as selenide and iodide, that are produced by the red blood cellsand released into the bloodstream and tissues is also reduced, resultingin a lower amount of reducing agents in the bloodstream and tissues,including, e.g., injured or diseased tissues. The lower amount ofreducing agents (iodide and selenide) results in a diminish capacity toneutralize free radicals, which result in oxidative stress and causestissue damage, e.g., damage resulting from hypoxia, ischemia, orreperfusion. This phenomenon is referred to as a “blood attack.” Thisand related mechanisms are discussed in, e.g., D'Alessandro, A. et al.Blood Transf, 2013: 11: 75-87; Salmi, H. et al., PediatricEndocrinology, Diabetes, and Metabolism 2011, 17, 1, 14-19; Snethil, S.et al., Clinica Chimica Acta 348 (2004) 131-137; and Tolan, N. et al.,Anal. Chem. 2009, 81, 31202-3108. Furthermore, addition of exogenousiodide and selenide into the blood stream relieves the burden on the redcells to produce selenide and iodide and enables the red cells to usethe reminding reducing power for the maintenance of hemoglobin.

Glutathione (GSH) is a tripeptide with a gamma peptide linkage betweenthe amine group of cysteine (which is attached by normal peptide linkageto a glycine) and the carboxyl group of the glutamate side-chain. It isan antioxidant, preventing damage to important cellular componentscaused by reactive oxygen species such as free radicals and peroxides.Thiol groups are reducing agents, existing at a concentration ofapproximately 5 mM in animal cells. Glutathione reduces disulfide bondsformed within cytoplasmic proteins to cysteines by serving as anelectron donor. In the process, glutathione is converted to its oxidizedform glutathione disulfide (GSSG), also called L-(−)-glutathione.Oxidized glutathione can be reduced by glutathione reductase, usingNADPH as an electron donor. The ratio of reduced glutathione to oxidizedglutathione within cells is often used as a measure of cellulartoxicity.

Accordingly, the present invention provides a method of treating orpreventing an injury or disease, including any of those describedherein, including but not limited to ischemic or reperfusion injury, ina subject in need thereof, by inhibiting or preventing a decrease in theratio of the amount or molar concentration of reduced forms ofglutathione to the amount or molar concentration of oxidized forms ofglutathione in the subject's bloodstream or at a site of disease orinjury, e.g., in a diseased or injured tissue. In certain embodiments,the method comprises providing to the subject an effective amount of acomposition of the present invention. In particular embodiments, thecomposition comprises an effective amount of one or more halides and/orone or more chalcogenides, and optionally one or more additional activeagents, including any of those described herein. The method may also beused to increase the ratio of the amount or molar concentration ofreduced forms of glutathione to the amount or molar concentration ofoxidized forms of glutathione in the subject's bloodstream or at thesite of disease or injury. In particular embodiments, the methodscomprise providing a halogen compound and/or a chalcogenide systemicallyto the subject prior to, at the time of, or after onset of said diseaseor injury. In certain embodiments, a halogen compound, e.g., iodide, andis provided to the subject prior to the onset of the disease or injury.In particular embodiments, a chalcogenide, e.g., selenide, is providedto the subject prior to the onset of the disease or injury, during theonset or occurrence of the disease or injury, of after the onset of thedisease or injury. It is further understood that other compounds thatimpede thyroid hormone production, activity or function, includinggoitrogens, may be used instead of or in combination with the halogencompound or chalcogenide to practice these or other methods of theinvention. Chalcogenides and halogen compounds may be used at any of thevarious amounts described herein. In certain embodiments, about 1 pg/kgto about 1 g/kg of chalcogenide, sulfide, or selenide is provided to thesubject, about 10 μg/kg to about 10 mg/kg of chalcogenide, sulfide orselenide is provided to the subject. In certain embodiments, the halogencompound is iodide and about 10 pg/kg to about 1 g/kg of iodide isprovided to the subject. In certain embodiments, about 10 μg/kg to about10 mg/kg of iodide is provided to the subject.

In certain embodiments, the methods of the invention inhibit or maintainthe ratio of the amount or molar concentration of reduced forms ofglutathione to the amount or molar concentration of oxidized forms ofglutathione in the subject's bloodstream or at the site of disease orinjury from falling below 99%, below 98%, below 97%, below 96%, below95%, below 94%, below 93%, below 92%, below 91%, below 90%, below 80%,below 70%, below 60%, below 50%, below 40%, below 30%, below 20%, orbelow 10% after onset of the disease or injury. In certain embodiments,the methods of the invention increase the ratio of the amount of reducedforms of glutathione to the amount of oxidized forms of glutathione inthe subject's bloodstream or at the site of disease or injury to at orabove 99%, at or above 98%, at or above 97%, at or above 96%, at orabove 95%, at or above 94%, at or above 93%, at or above 92%, at orabove 91%, at or above 90%, at or above 80%, at or above 70%, at orabove 60%, at or above 50%, at or above 40%, at or above 30%, at orabove 20%, or at or above 10% after onset of the disease or injury. Inparticular embodiments, the recited ratios are achieved within ormaintained for 1 minute, 5 minutes, 30 minutes, one hour, 2 hours, fourhours, eight hours, 12 hours, 24 hours, 48 hours, or 48 hours of thetime when the composition is provided to the subject.

The present invention further provides methods of determining ormonitoring the therapeutic effect of a composition or treatment,including any of the compositions or methods described herein, which maybe practiced alone or in combination with any of the methods describedherein. These methods comprise determining the ratio of the amount orconcentration of reduced glutathione to the amount or molarconcentration of oxidized glutathione. The ratio of the amount or molarconcentration of reduced glutathione to the amount or molarconcentration of oxidized glutathione may be determined, e.g., for ablood sample, e.g., as described in any of the previously citedarticles. An increase or inhibition of further reduction in the ratiofollowing treatment is indicative of efficacy, whereas a decrease in theratio suggests that the treatment has limited or no therapeutic value.

In some embodiments, the present invention relates to a method ofpreventing, inhibiting, or reducing an immune response in a biologicalmaterial or subject in need thereof, comprising providing to thebiological material or subject an effective amount of a composition ofthe present invention. In particular embodiments, the compositioncomprises an effective amount of one or more halides and/or one or morechalcogenides, and optionally one or more additional active agents,including any of those described herein. In some embodiments, an immuneresponse generally refers to any response elicited by the innate oradaptive immune system. In some embodiments, the immune response isinitiated by the activation of a T cell. In some embodiments, the immuneresponse is initiated by the activation of a B cell. Immune responsescontemplated herein may be stimulated any means. In some embodiments, animmune response is induced by a pathogen. In some embodiments, an immuneresponse is an autoimmune response. In some embodiments, immune responserefers to inflammation. In certain embodiments, an autoimmune responseis induced following transplantation of cells, tissue or an organ. Incertain embodiments, an autoimmune response is the result of anautoimmune disorder.

In some embodiments, inflammation refers to a non-specific firstreaction mounted by the immune system in response to a perceived injuryor threat. It is an innate defensive response, distinguished from themore precisely tailored adaptive responses of the immune system.Inflammation may work cooperatively with adaptive responses of theimmune system, which develop more slowly but are more precisely targetedto a harmful agent such as a chemical or pathogen that may be causinglocalized injury.

Inflammation may be associated with infections, but it occurs inresponse to virtually any type of injury or threat, including physicaltrauma, cold, burns from radiation, heat or corrosive materials,chemical irritants, bacterial or viral pathogens, localized oxygendeprivation (ischemia) or reperfusion (sudden reinfusion of oxygen toischemic tissue), and others. It includes the classic symptoms ofredness, heat, swelling, and pain, and may be accompanied by decreasedfunction of the inflamed organ or tissue. It may be a generalizedreaction involving several effects that may tend to combat an injuriousagent that may be present at the site where an injury or threat wasdetected, or it may tend to contain the injury or threat to its initiallocation, to keep it from spreading rapidly.

In some embodiments, an autoimmune response is generally defined as anyresponse in an organism that results in a immune attack against its owncells and tissues. The etiology of autoimmunity is very broad,comprising genetic and environmental factors as well as immunologicalmechanisms including, e.g, but not to be limited in anyway: T-CellBypass; T-Cell-B-Cell discordance; Aberrant B cell receptor-mediatedfeedback; Molecular Mimicry; Idiotype Cross-Reaction; CytokineDysregulation; Dendritic cell apoptosis; Epitope spreading, epitopedrift; Epitope modification or Cryptic epitope exposure.

In some embodiments, autoimmunity refers to systemic autoimmunediseases, which are defined as autoimmune diseases that damage manyorgans; such as e.g., lupus erythematosus, Sjögren's syndrome,scleroderma, rheumatoid arthritis, and autoimmune myositis such asdermatomyositis, polymyositis, vasculitis (e.g., Churg-StraussSyndrome), Sarcoidosis, general allergies etc.

In some embodiments, autoimmunity refers to local autoimmune syndromes,which are defined as autoimmune disease that affect a single organ ortissue; such as e.g., diabetes mellitus type 1; Hashimoto's thyroiditis;Addison's disease; Celiac disease; Crohn's Disease; Graves' disease,Pernicious anaemia; Psoriasis; Pemphigus vulgaris; Vitiligo; Autoimmunehaemolytic anaemia; Idiopathic thrombocytopenic purpura; Myastheniagravis; etc.

Some embodiments of the present invention relate to the treatment,prevention, or amelioration of organ rejection or graft-versus-hostdisease (GVHD) and/or conditions associated therewith. In certainembodiments, the present invention includes methods of treating orpreventing organ rejection or graft-versus-host disease (GVHD) and/orconditions associated therewith in a subject in need thereof, comprisingproviding to the subject an effective amount of a composition of thepresent invention. In certain embodiments, the present inventionincludes methods of treating or preventing organ rejection orgraft-versus-host disease (GVHD) and/or conditions associated therewithin a subject in need thereof, comprising providing contacting thetransplanted cells, tissue or organ with an effective amount of acomposition of the present invention. In particular embodiments, thecomposition comprises an effective amount of one or more halides and/orone or more chalcogenides, and optionally one or more additional activeagents, including any of those described herein. Organ rejection occursby host immune cell destruction of the transplanted tissue through animmune response. Similarly, an immune response is also involved in GVHD,but, in this case, the foreign transplanted immune cells destroy thehost tissues. The administration of the halogen or chalcogenidecompounds of the invention, that inhibit an immune response, may be aneffective therapy in preventing organ rejection or GVHD.

Some embodiments relate to the treatment, prevention, or amelioration ofinflammation resulting from an ischemic or reperfusion injury. Incertain embodiments, the present invention includes methods of treatingor preventing inflammation, e.g., inflammation resulting from anischemic or reperfusion injury in a subject in need thereof, comprisingproviding to the subject an effective amount of a composition of thepresent invention. In certain embodiment, the composition comprises aneffective amount of a halide and/or an effective amount of achalcogenide, and optionally one or more additional active agents,including any of those described herein. In certain embodiments,reperfusion injuries occur when blood supply to an organ has been cutoff for a period of time, such as e.g. following myocardial infarction,stroke, and other thrombotic events. During ischemia, tissues becomestressed due to lack of oxygen, and eventually, if the ischemia isprolonged, the tissue will become necrotic, resulting in theaccumulation of various metabolic intermediates. Restoration of bloodflow results in a rapid increase in oxygen radical that lead toinflammation. Thus embodiments of the present invention relate to theuse of halogen compounds such as iodide, or chalcogenide compounds suchas selenide or sulfide to reduce the formation of the reactive oxygenspecies.

There are many different assays described in the art that are useful fordetecting the activation of an immunological response. Some typicalnon-limiting examples of these include, but are in know way limited to:methods to detect lymphocytes (i.e., T cells, B cells, and NK cells),monocytes, and dendritic cells. Examples of such assays include e.g.,ELISPOT; flow cytometry-based analysis of cytokine expression;HLA-peptide multimer staining; MHC Tetramer analysis; carboxyfluoresceinsuccinimidyl ester assay; antigen-specific immune responses:delayed-type hypersensitivity, PCR-based detection of T-cell receptorgene usage or cytokine production, mixed lymphocyte reactions to measureT cell proliferation, etc.

In embodiments, any assay capable of detecting an immune responsegenerated by any stimulus, is useful for measuring the effect that ahalogen compound or a chalcogenide compound may have on the suppressionof an immune response.

In particular embodiments, immunosuppression induced by halogen orchalcogenide compounds is assessed by measuring T cell proliferation,e.g., in response to activated antigen presenting cells. Essentially amixed lymphocyte reaction, this assay exploits that fact that T cellexposure to presented antigens result in T cell activation andproliferation, as well as supportive increases in DNA synthesis. PBMCswere isolated from two donors and then the samples were split into twogroups, one that receives irradiation and one that does not. Irradiationresults in the presentation of numerous antigens to antigen presentingcells, thus the irradiated cells serve as potent activators of T cells.By mixing the irradiated cells with the non-irradiated cells, strong Tcell proliferation will occur. This proliferation can be measured usingany proliferation assay, as would be commonly known to the skilledartisan. In one embodiment, proliferation is assessed by measuring DNAsynthesis. Mixed PBMC were cultured in the presence or absence ofhalogen or chalcogenide compounds for 5 days, and then pulsed withtritiated thymidine for 18 hours to label newly synthesized DNA. Thedegree of labeling is directly correlated with the degree of T cellactivation, thus low levels of labeling correlate withimmunosuppression.

In some embodiments, halogen or chalcogenide compound immunosuppressionis assessed using this assay, and the halogen compound is found toresult in a 10% reduction in tritiated thymidine incorporation, thusindicating 10% immunosuppression. In some embodiments, treatment with ahalogen compound in this way results in 20% immunosuppression, or 30%,40%, 50%, 60%, 70%, 80%, 90%, or 100% immunosuppression. In particularembodiments, the halogen compound comprises iodide. In particularembodiments the chalcogenide compound that induces immunosuppressioncomprises selenide or sulfide.

In certain embodiments of methods of the invention, the amount of oreffective amount of a halogen compound that is provided to a biologicalmaterial, e.g., a mammal or tissue therein, is about, at least, at leastabout, or at most about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39,40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150, 160, 170, 180,190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320,330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 441, 450,460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590,600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730,740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870,880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, 1000 mg,mg/kg, or mg/m², or any range derivable therein. Alternatively, theamount may be expressed as 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240,250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380,390, 400, 410, 420, 430, 440, 441, 450, 460, 470, 480, 490, 500, 510,520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650,660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790,800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930,940, 950, 960, 970, 980, 990, 1000 mM or M, or any range derivabletherein.

In certain embodiments of methods of the invention, the halogen compoundcomprises iodine or iodide, e.g., NaI, and the effective amount isgreater than or equal to about 150 μg, greater than or equal to about300 μg, greater than or equal to about 500 μg, greater than or equal toabout 1 mg, greater than or equal to about 2 mg, greater than or equalto about 5 mg, greater than or equal to about 10 mg, greater than orequal to about 15 mg, or greater than or equal to about 20 mg. Incertain embodiments, the effective amount is 150 μg to 1000 mg, 300 μgto 1000 mg, 500 μg to 1000 mg, 1 mg to 1000 mg, 2 mg to 1000 mg, 5 mg to1000 mg, 10 mg to 1000 mg, 150 μg to 100 mg, 300 μg to 100 mg, 500 μg to100 mg, 1 mg to 100 mg, 2 mg to 100 mg, 5 mg to 100 mg, or 10 mg to 100mg. In certain embodiments, the effective amount is 150 μg to 50 mg, 300μg to 20 mg, 500 μg to 10 mg, 1 mg to 20 mg, 1 mg to 10 mg, or about 5mg, about 10 mg, about 15 mg, or about 20 mg.

In particular embodiments of any of the methods of the presentinvention, a biological material, e.g., a subject, is treated with orcontacted with an effective amount of a composition or compound of thepresent invention, wherein said effective amount of about 0.01 mg/kg toabout 20 mg/kg, about 0.05 mg/kg to about 10 mg/kg, about 0.1 mg/kg toabout 5 mg/kg, about 0.5 mg/kg to about 2 mg/kg, about 0.5 mg/kg toabout 1 mg/kg, about 0.5 mg/kg, about 0.6 mg/kg, about 0.7 mg/kg, about0.8 mg/kg, about 0.9 mg/kg, about 1.0 mg/kg, about 1.1 mg/kg or about1.2 mg/kg.

In particular embodiments, an effective amount of iodine or iodide is anamount at least or about two-fold, three-fold, four-fold, five-fold,six-fold, seven-fold, eight-fold, nine-fold, ten-fold, twelve-fold,fifteen-fold, or twenty-fold of the average daily recommended amounts aslisted below. In particular embodiments, the effective amount of iodineor iodide is an amount between two-fold and twenty-fold, betweenfive-fold and fifteen-fold, or between five-fold and ten-fold of theaverage daily recommended amounts of iodine as listed below.

Recommended Amount¹ Life Stage (mcg) Birth to 6 months 110 Infants 7-12months 130 Children 1-8 years 90 Children 9-13 years 120 Teens 14-18years 150 Adults 150 Pregnant teens and women 220 Breastfeeding teensand women 290 ¹NIH Office of Dietary Supplements Iodine Fact Sheet forConsumers, reviewed Jun. 24, 2011, obtained 2013.

In particular embodiments of any of the methods of the presentinvention, the methods further comprising measuring or determining theamount or concentration of halogen compound present in a biologicalmaterial or subject to be treated (e.g., the concentration present in asubject's bloodstream), and then providing to the biological material orsubject an amount of composition comprising a halogen compound toachieve a desired total amount or concentration of halogen compound inthe biological material or subject, wherein the total amount orconcentration includes both the amount or concentration measured ordetermined in the biological material or subject and the amount providedto the biological material or subject. In particular embodiments, thedesired total amount is an effective amount for the treatment orprevention of the disease or injury afflicting a subject, or aneffective amount for preventing inflammation or rejection of atransplanted organ. In certain embodiments of methods of the invention,the halogen compound comprises iodine, e.g., NaI, and the effectiveamount is about 0.01 mg/kg to about 20 mg/kg, about 0.05 mg/kg to about10 mg/kg, about 0.1 mg/kg to about 5 mg/kg, about 0.5 mg/kg to about 2mg/kg, about 0.5 mg/kg to about 1 mg/kg, about 0.5 mg/kg, about 0.6mg/kg, about 0.7 mg/kg, about 0.8 mg/kg, about 0.9 mg/kg, about 1.0mg/kg, about 1.1 mg/kg or about 1.2 mg/kg. In certain embodiments, thehalogen compound comprises iodine, and the effective amount is an amountthat achieves about the same concentration or amount that is achieved byan effective amount of iodine that is at least or about two-fold,three-fold, four-fold, five-fold, six-fold, seven-fold, eight-fold,nine-fold, ten-fold, twelve-fold, fifteen-fold, or twenty-fold of theaverage daily recommended amounts as listed below. In particularembodiments, the effective amount of iodine or iodide is an amountbetween two-fold and twenty-fold, between five-fold and fifteen-fold, orbetween five-fold and ten-fold of the average daily recommended amountsof iodine as listed below.

Recommended Amount¹ Life Stage (mcg) Birth to 6 months 110 Infants 7-12months 130 Children 1-8 years 90 Children 9-13 years 120 Teens 14-18years 150 Adults 150 Pregnant teens and women 220 Breastfeeding teensand women 290 ¹NIH Office of Dietary Supplements Iodine Fact Sheet forConsumers, reviewed Jun. 24, 2011, obtained 2013.

In additional embodiments wherein a composition comprising a halogencompound is provided in combination with one or more additional activeagent (or composition comprising the same), the amount or effectiveamount of the additional active agent is any of the concentrations ordosages described herein with respect to the halogen compound. Inaddition, the additional active agent may be provided by any of theroutes of administration described herein with respect to thecomposition comprising the halogen compound. In addition, thecomposition comprising the halogen compound and the compositioncomprising the additional active agent, e.g., a chalcogen such as asulfide or selenide, may be provided to the subject at the same time, atdifferent times, or during overlapping time periods.

In particular embodiments where the composition comprises glutathioneand either or both of a halogen compound and a chalcogenide, the halogencompound and chalcogenide (when present) are administered at any of theamounts described herein for either active agent. In certain embodimentsglutathione is present in an amount sufficient to inhibit oxidation ofthe chalcogenide and/or halogen compound, including any of the rangesdescribed herein. In particular embodiments, the halogen compound isiodide, e.g., NaI, and the chalcogenide is sulfide or selenide.

In particular embodiments of the present invention, an effective amountof a composition is administered to a biological material, e.g., atissue or subject, prior to reperfusion, e.g., for at least one, twothree, four, five or ten minutes of the ten minutes or thirty minutesimmediately prior to ischemia or reperfusion. In particular embodiments,this administration of the composition results in reduced ischemic orreperfusion injury. In certain embodiments, the reperfusion injury isless than 80%, less than 70%, less than 60%, less than 50%, less than40%, or less than 30% the reperfusion injury in the absence of treatmentwith the composition.

In particular embodiments, treatment of a biological material, i.e., asubject, with a composition of the present invention following aninjury, e.g., heart attack or stroke, results in a reduced injury, e.g.,infarct size. In certain embodiments, the injury or infarct size is lessthan 80%, less than 70%, less than 60%, less than 50%, less than 40%, orless than 30% the severity or infarct size in the absence of treatmentwith the stable composition.

In particular embodiments, treatment of a biological matter, e.g., asubject within 10 minutes, within 30 minutes, within one hour, or withintwo hours or a heart attack, with a composition of the present inventionresults in a decrease in heart damage or an increase in fractionalshortening or left ventricular function, as compared to in the absenceof treatment with the composition. In particular embodiments, thedecrease in heart damage is at least 10%, at least 20%, at least 30%, atleast 40%, or at least 50%, as compared to in the absence of treatmentwith the composition. Decrease in heart damage may be detected byassessing levels of the heart specific protein, cardiac troponin I, inthe blood of a subject after treatment. In particular embodiments, theincrease in fractional shortening or left ventricular function is atleast 20%, at least 30%, at least 40%, or at least 50%, as compared toin the absence of treatment with the composition.

In certain embodiments, the present invention includes a method ofreducing inflammation in the heart of a subject, e.g., a mammal, afterheart attack, by administering to the subject a composition of thepresent invention. In certain embodiments, the decrease in heartinflammation is at least 10%, at least 20%, at least 30%, at least 40%,or at least 50%, as compared to in the absence of treatment with thecomposition.

In various embodiments of methods of the present invention, a biologicalmaterial, e.g., an organ, a subject, or a tissue therein, is exposed toa composition of the current invention for about, at least, at leastabout, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30,35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7days or more, and any range or combination therein.

Furthermore, when administration of a composition according to thepresent invention is intravenous or by infusion, it is contemplated thatthe following parameters may be applied. A flow rate of about, at leastabout, or at most about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50,51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 gtts/min orμgtts/min, or any range derivable therein. In some embodiments, theamount of the composition is specified by volume, depending on theconcentration of the halogen compound present in the composition. Anamount of time may be about, at least about, or at most about 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58,59, 60 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days, 1, 2, 3,4, 5 weeks, and/or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, or anyrange derivable therein.

Volumes of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53,54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150,160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290,300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430,440, 441, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560,570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700,710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840,850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980,990, 1000 mls or liters, or any range therein, may be administeredoverall or in a single session.

According to various embodiments of the methods of the presentinvention, a biological material is provided with a composition of theinvention, e.g., intravenously, intradermally, intraarterially,intraperitoneally, intralesionally, intracranially, intraarticularly,intraprostaticaly, intrapleurally, intratracheally, intranasally,intravitreally, intravaginally, intrarectally, topically,intratumorally, intramuscularly, intraperitoneally, intraocularly,subcutaneously, subconjunctival, intravesicularly, mucosally,intrapericardially, intraumbilically, intraocularally, orally,topically, locally, by injection, by infusion, by continuous infusion,by absorption, by adsorption, by immersion, by localized perfusion, viaa catheter, or via a lavage. In particular embodiments, it is providedparenterally, e.g., intravenously, or by inhalation. “Parenteral” refersto any route of administration of a substance other than via thedigestive tract. In specific embodiments, a halogen compound is providedto the subject by intravenous administration or infusion.

In certain embodiments, a method or composition of the present inventionis used to treat or prevent a disease or injury selected from any of thefollowing, or which is caused by or results from any of the following:ablation therapy, adrenalectomy, aortic aneurysm, aortic root surgery,aortic stenosis, aortic valve disease, arrhythmia, atherosclerosis,atrial flutter, atrial fibrillation, atrial septal defect, arteriovenousmalformation, awake brain surgery, bariatric surgery, bone marrowtransplant, brachial plexus injuries, bradycardia, brain aneuryism,breast augmentation surgery, breast reduction surgery, burn injury,coronary bypass surgery, coronary artery disease, cardiac ablation,cardiac catheterization, cardiac resynchronization therapy, cardiacsurgery, cardiomyopathy, cardiac surgery, cardiovascular diseases,carotid angioplasty and stenting, coarctation of the aorta, congenitalheart disease, coronary bypass surgery, coronary artery disease,critical care medicine, chronic obstructive pulmonary disease, elbowreplacement surgery, emergency medicine, general internal medicine,general surgery, gastrointestinal bleeding, heart attack, hearttransplant, heart valve surgery, hip replacement surgery, hypertrophiccardiomyopathy, hypoxia ischemia encephalopathy, hysterectomy, ileoanalanastomosis (j pouch) surgery, inflammatory bowel disease, ischemicheart disease, ischemia reperfusion injury, irritable bowel syndrome,jaw surgery, kidney transplant, laryngotracheal reconstruction surgery,liver transplant, lung volume reduction surgery, lung transplant,minimally invasive heart surgery, neurosurgery, oral and maxillofacialsurgery, orthopedic surgery, pancreas transplant, pancreatitis, partialnephrectomy, pediatric cervical spine surgery, pediatric surgery, pelvicorgan prolapse, plastic and reconstructive surgery, pulmonary andcritical care medicine, pulmonary atresia, pulmonary vein isolation,rectal prolapse, restrictive cardiomyopathy, retinal detachment,retinopathy of prematurity, robotic surgery, spinal cord injury,spontaneous coronary artery dissection, spontaneous occlusion of thecircle of willis, stroke, stroke telemedicine (telestroke), suddencardiac arrest, stereotactic radiosurgery, surgery, chronic heartfailure, systolic heart failure, tachycardia, teare's disease, thoracicaortic aneurysm, thoracic surgery, total elbow arthroplasty, tricuspidvalve disease, ulcerative colitis, valve-preserving aortic root repair,vascular and endovascular surgery, vascular medicine, or ventriculartachycardia.

In certain embodiments, the injury or disease is selected from, resultsfrom or is caused by inflammation, heart attack, coronary bypasssurgery, ischemia, gut ischemia, liver ischemia, kidney ischemia,hypoxic-ischemic encephalopathy, stroke, traumatic brain injury, limbischemia, eye ischemia, sepsis, smoke, burn, reperfusion, or acute lunginjury. In certain embodiments, the injury is an infarct caused by aheart attack or a stroke. In particular embodiments, the injury iscaused by coronary bypass surgery, optionally a coronary artery bypassgraft (CABG). In one embodiment, the disease is hypoxic-ischemicencephalopathy.

Delivery Devices

In additional embodiments, the present invention includes a drugdelivery device designed to limit, prevent or inhibit oxidation of areduced form of an active agent, such as, e.g., a reduced form of ahalogen compound or a reduced form of a chalcogenide. In specificembodiments, the device maintains a reduced form of an active agent inits reduced form. In particular embodiments, the device comprises thereduced form of an active agent, such as the reduced form of a halogencompound or chalcogenide, for example. In specific embodiments, the drugdevice comprises a composition of the present invention.

Manufacturer-prepared, premixed ready-to-use products represent a usefulapproach to intravenous drug safety, since they remove error associatedwith measuring and diluting intravenous or infused drugs. Accordingly,in certain embodiments, the present invention includes a drug deliverydevice for administration of a ready-to-use product comprising a reducedform of an active agent. In particular embodiments, the reduced form ofactive agent is a reduced form of a halogen compound and/or achalcogenide, e.g., selenide or sulfide.

In related embodiments, the present invention comprises a containerhaving therein an effective amount of a composition of the presentinvention or an effective amount of a halogen compound. The effectiveamount may be in liquid form, e.g., the active agent may be dissolved ina solution, or it may be in dry form (e.g., dried, lyophilized, orfreeze-dried), such that the active agent may be dissolved in a solutionprior to administration to a subject.

In all embodiments of compositions described herein, it is understoodthat the composition may be a pharmaceutical composition.

In addition, it is understood that traditional methods for deliveringactive agents, which can involve injecting air into a vial comprisingthe active agent during the process of withdrawing the active agent intoa syringe or bag, may result in the undesired oxidation of an activeagent that is in a reduced form. Accordingly, the present inventioncontemplates delivery devices that minimize of prevent contact of anactive agent with oxygen during delivery to a subject.

In one embodiment, the present invention includes a drug deliverydevice, comprising:

a reservoir for containing a composition of the present invention, i.e.,comprising a halogen compound or reduced form of a halogen compound,alone or in combination with one or more additional active agents, suchas, e.g., a chalcogenide or a reduced form of a chalcogenide, such as asulfide or selenide; and

a fluid communicator, the fluid communicator configured to maintain atleast 90% of the reduced form of active agent in the composition inreduced form during delivery to a subject. In particular embodiments, itis configured to maintain at least 95%, at least 96%, at least 97%, atleast 98%, or at least 99% of the reduced form of active agent inreduced form during administration to a subject. In certain embodiments,the fluid communicator is in fluid communication with the reservoir. Inother embodiments, it is can be placed into fluid communication with thereservoir.

In certain embodiments, the reservoir is oxygen impermeable, and/or itis formed of an oxygen impermeable polymer. In certain embodiments, thereservoir comprises an oxygen impermeable layer, e.g., at its innersurface, which may comprise: polyethylene (LDPE), polystyrene (PS),polyethylene (PE), polycarbonate (PC), polyvinyl acetate (PVAc), APET,polyvinly chloride (PVC), nylon 6 (Ny6), polyvinyl fluoride (PVF),polyvinylidene chloride (PVdC), polyacetonitrile (PAN), ethylene vinylalcohol (EVOH), or Polyvinyl alcohol (PVA). In certain embodiments, theoxygen transmission coefficient of said polymer is less than 10⁻¹⁰[cm³(STP)/cm/(cm²+s+Pa)]. In related embodiments, it is less than 10⁻⁹,less than 10⁻⁸, or less than 10⁻⁷ [cm³(STP)/cm/(cm²+s+Pa)]. In relatedembodiments, the reservoir includes multiple layers of oxygenimpermeable polymers.

In particular embodiments, the reservoir includes a resealable port. Aresealable port may be used for introducing a solution comprising atherapeutic agent, such as a reduced form of a halogen compound and/or areduced form of a chalcogenide, into the reservoir. A resealable portmay be used for coupling to the fluid communicator. In particularembodiments, the reservoir includes a plurality of resealable ports. Incertain embodiments, the reservoir is a bottle, a bag, a tube, a vial,or a syringe. In particular embodiments, it is an intravenous bag or asyringe. In particular embodiments, it is a tubular member, the tubularmember having a septum configured to hermetically seal the tubularmember. In one embodiment, the tubular member is a Hungate tube.

In particular embodiments, the fluid communicator is configured to befluidically coupled to the reservoir through a resealable port. Inrelated embodiments, the fluid communicator is configured tointravenously deliver an active agent, e.g., a reduced form of a halogencompound and/or a reduced form of a chalcogenide, from the reservoir toa subject in need thereof. In certain embodiment, the fluid communicatorincludes at least one of a needle and a cannula.

In certain embodiments, the device, including the fluid communicator, isdisposed in a reduced oxygen or oxygen-free environment. In particularembodiments, the reduced oxygen or oxygen-free environment is within acontainer, optionally wherein said container is a bag or a malleablecontainer, e.g., that allows manipulation of components of the devicethrough the bag, such as coupling of the fluid communicator to thereservoir in embodiments where they are not initially coupled. Inparticular embodiment, the container comprises one or more oxygenimpermeable polymer, including any of those described above. Inparticular embodiments, the walls of the container comprise multiplelayers of one or more oxygen impermeable polymers.

In particular embodiments, the device comprises in the reservoir acomposition comprising a reduced form of a halogen compound and/or achalcogenide. In certain embodiments, the reduced form of halogencompound is an iodide, e.g., sodium iodide, potassium iodide, orhydrogen iodide, and/or the reduced form a chalcogenide is H₂Se, Na₂Seor NaHSe or a sulfide compound.

In particular embodiments, the device allows for delivery of the reducedform of active agent, e.g., a reduced form of a halogen compound, suchas iodide and/or a reduced form of a chalcogenide, such as selenide orsulfide, wherein at least 90%, at least 95%, at least 96%, at least 97%,at least 98%, or at least 99% of the active agent is delivered to thesubject in reduced form.

The present invention further includes a method for reducing injury ortreating a disease in a subject by providing to said subject acomposition of the invention, wherein said composition is provided tosaid subject using the device of the present invention. In particularembodiments, the injury or disease is any of those described herein,including but not limited to an injury resulting from ischemia orreperfusion. In particular embodiments, the injury is an infarct causedby a heart attack or stroke, or chronic hear failure. In otherembodiments, the injury is caused by inflammation. In certainembodiments, the composition comprising a halogen compound is providedto a subject using a device described herein. In certain embodimentswherein the composition comprising a halogen compound also comprises anadditional therapeutic agent or wherein the composition comprising ahalogen compound is provided in combination with a compositioncomprising an additional active agent, such as a reduced form of achalcogenide, the composition comprising the halogen compound and/or theother composition are provided using a device described herein.

Further, the present invention also includes a method of delivering acomposition comprising a reduced form of an active agent, wherein thecomposition comprises a halogen compound, to a patient in need thereof,the method comprising:

containing the composition comprising a reduced form of active agent ina reservoir, the reservoir configured to maintain the active agent in areduced form;

establishing fluid communication between the reservoir and the subject;

delivering a predetermined volume of the composition from the reservoirto the subject in an environment substantially free of oxygen.

In particular embodiments, a device of the present invention comprises:(1) an i.v. bag comprising a composition of the present invention,wherein said i.v. bag is oxygen impermeable; and (2) a tubing, whereinsaid tubing is coupled at a first end of the tubing to the i.v. bagthrough a port in the i.v. bag, or wherein said tubing is capable ofbeing coupled at a first end of the tubing to the i.v. bag through aport in the i.v. bag, wherein said tubing is coupled at a second end ofthe tubing to a needle or cannula, or wherein said tubing is capable ofbeing coupled at a second end of the tubing to a needle or cannula,wherein said device is contained within a bag comprising a reducedoxygen or oxygen-free environment. In particular embodiments, the i.v.bag and/or the bag comprise one or more oxygen impermeable polymersdescribed herein. In particular embodiments, the bag is flexible, suchas to allow a user to couple the tubing to the i.v. bag and/or needlewhile the device remains sealed in the bag.

In particular embodiments, a device of the present invention comprises:(1) an i.v. bag comprising a composition of the present invention,wherein said i.v. bag is oxygen impermeable; and (2) a tubing, whereinsaid tubing is coupled at a first end of the tubing to the i.v. bagthrough a port in the i.v. bag, or wherein said tubing is capable ofbeing coupled at a first end of the tubing to the i.v. bag through aport in the i.v. bag, wherein said tubing is coupled at a second end ofthe tubing to a needle or cannula, or wherein said tubing is capable ofbeing coupled at a second end of the tubing to a needle or cannula,wherein said device is contained within a bag comprising a reducedoxygen or oxygen-free environment. In particular embodiments, the i.v.bag and/or the bag comprise one or more oxygen impermeable polymersdescribed herein. In particular embodiments, the bag is flexible, suchas to allow a user to couple the tubing to the i.v. bag and/or needlewhile the device remains sealed in the bag, and/to apply pressure to thei.v. bag to begin flow of the composition through the tubing towards orinto the needle or cannula.

In particular embodiments, a device of the present invention comprises:(1) an syringe comprising a composition of the present invention,wherein said syringe is oxygen impermeable; and (2) a needle or cannula,wherein said needle or cannula is coupled to the syringe through a portin the syringe, or wherein said needle or cannula is capable of beingcoupled to the syringe through a port in the syringe, wherein saiddevice is contained within a bag comprising a reduced oxygen oroxygen-free environment. In particular embodiments, the syringe and/orthe bag comprise one or more oxygen impermeable polymers describedherein. In particular embodiments, the bag is flexible, such as to allowa user to couple the needle or cannula to the syringe while the deviceremains sealed in the bag, and/to apply pressure on the syringe to beginflow of the stable composition into the needle or cannula.

In particular embodiments, the device comprises a therapeuticallyeffective amount of an agent or composition of the present invention.

EXAMPLES Example 1 Sodium Iodide Improves Myocardial Infarction Outcome

This example demonstrates that sodium iodide (NaI) improves outcome in amouse model of heart attack by reducing infarct size.

In order to examine the effect of NaI on myocardial infarct size in amouse model of heart attack, open chest ligation of the left descendingcoronary artery (LAD ligation) was performed in mice. At 55 minutes postinitiation of ischemia, either vehicle (saline) or test agent (NaI) wasinjected into the femoral vein of each mouse. The heart muscle wasreperfused by releasing the ligature at 60 minutes post initiation ofischemia. After 2 hours of reperfusion, infarct size was measured bymorphometry (FIG. 1). Five groups of mice were utilized in this study,including a saline vehicle group (n=3) and five test groups thatreceived 0.25 mg/kg (n=4), 0.5 mg/kg (n=5), 1 mg/kg (n=5), or 2 mg/kg(n=6) of NaI. To be certain that all animals in the study were treatedin a similar way, the area at risk (AAR) as a proportion of the leftventricle was determined (FIG. 1). Area at risk (AAR) and infarct size(Inf) were measured using the specific dyes TTC and Evans blue.Processed images were generated in an unbiased way by setting constantthresholds and allowing Photoshop® to generate the images that werequantitated to generate the data presented in FIG. 1.

These results show that there were statistically significant decreasesin infarct size when comparing the saline group to all four test groupsthat received NaI (FIG. 1). For each dosage tested, treatment with NaIresulted in an approximately 70% reduction in infarct size in comparisonto vehicle alone.

An additional test was performed under the same conditions but using anexpanded range of concentration of NaI. The results of this test alsoshow that there were statistically significant decreases in infarct sizewhen comparing the saline group to all four test groups that receivedNaI (FIG. 2).

To determine whether pretreatment with NaI prior to heart attackimproves heart attack outcome, open chest ligation experiments wereperformed in mice where either vehicle (saline) or test agent (NaI) wasinjected into the femoral vein of each mouse either 5 minutes or 48hours before initiation of ischemia. Open chest ligation of the leftdescending coronary artery (LAD ligation) was then performed, and theheart muscle was reperfused by releasing the ligature at 60 minutes postinitiation of ischemia. After 2 hours of reperfusion, infarct size wasmeasured by morphometry as described above (FIG. 3). Three groups ofmice were utilized in this study, including a saline vehicle group (n=1)and two test groups that received 1 mg/kg NaI 5 minutes before LADligation (n=4) or 1 mg/kg NaI 48 hours before LAD ligation (n=1).

These results show that there were statistically significant decreasesin infarct size when comparing the saline group to both test groups thatreceived NaI (FIG. 3). For each dosage tested, pre-treatment with NaIresulted in an approximately 70% reduction in infarct size in comparisonto vehicle alone.

In addition, the protective effect of NAI was confirmed byechocardiography, as shown in FIG. 4. In vivo transthoracicechocardiography of the left ventricle (LV) using a 30-MHz RMV scanheadinterfaced with a Vevo 770 (Visualsonics) was performed, andtwo-dimensional echocardiography images were obtained. LV end-diastolicdiameter (EDD) and LV end-systolic diameter (ESD) were measured at a daybefore for a baseline and one and 4 weeks after myocardial ischemia andreperfusion. LV percent fractional shortening (FS) and LV ejectionfraction (EF) were calculated.

Example 2 Glutathione Stabilizes Selenide

To demonstrate that glutathione prevents selenide from oxidation,solutions of 50 mM selenide were prepared in either water or 150 mM GSHand observed for the 8 minutes immediately following preparation. InFIG. 5, the oxidized forms of selenide appear dark in the solutions,with the samples in glutathione clearly showing reduced levels ofoxidation at each time point.

Example 3 Immunosuppressant Properties of Halogen Compounds andChalcogenides

To demonstrate the immunosuppressant properties of iodide and selenide,blood was obtained from two donors (A and B), and peripheral bloodmononuclear cells (PBMC) were purified from the blood. PBMC were eitherirradiated with 3500 rads (Ax and Bx) or not (A and B). Irradiated andunirradiated PBMC from each patient were mixed (100,000 from each) andplaced into culture medium containing various amounts of cyclosporin A(CSA, positive control), iodide (Iod), selenide (Sel), or just the mediafor 5 days. The specific amounts of each compound are shown in FIG. 6.Following this incubation, the cells were pulsed with tritiatedthymidine for 18 hours to determine the amount of DNA synthesis. Cellswere lysed, the lysate filtered to trap the DNA on the filter and thefilters counted in a scintillation counter. The amounts of incorporatedtritium were recorded and are shown in FIG. 7. PBMC from donor A notmixed with others cells or mixed with irradiated PBMC from donor Aserved as negative controls; likewise for donor B. The positive controlwas PBMC from A mixed with Bx or visa versa. All concentrations ofselenide reduced the labeling to control levels. Iodide at the highestconcentration reduced the amount of labeling. Others have shown that thedegree of labeling is directly correlated with the degree of T cellactivation. Low levels of labeling correlate with immunosuppression.These experiments demonstrate that selenide and iodide haveimmunosuppressant effects.

Example 4 Sodium Iodide Improves Myocardial Infarct Outcome

This example demonstrates that treatment with sodium iodide (NaI) indrinking water for two days prior to induced infarct improves outcome ina mouse model of heart attack.

In order to examine the effect of pre-treatment with drinking watercontaining NaI on myocardial infarct size in a mouse model of heartattack, three groups of mice were provided with drinking watercontaining various concentrations of iodide to achieve dosages of 0.07mg/kg/day (normal drinking water) for the first group; 0.77 mg/kg/dayfor the second group; or 7.07 mg/kg/day for the third group, for twodays immediately prior to open chest ligation of the left descendingcoronary artery (LAD ligation) to initiate ischemia. A fourth group ofmice was provided with normal drinking water for the two days prior tothe initiation of ischemia, but at 55 minutes post initiation ofischemia, either vehicle (saline) or test agent (NaI) was injected intothe femoral vein of each of these mice. The heart muscles of the micewere then reperfused by releasing the ligatures at 60 minutes postinitiation of ischemia. After two hours of reperfusion, infarct size wasmeasured by morphometry (FIG. 1). To be certain that all animals in thestudy were treated in a similar way, the area at risk (AAR) as aproportion of the left ventricle was determined (FIG. 8). Area at risk(AAR) and infarct size (Inf) were measured using the specific dyes TTCand Evans blue. Processed images were generated in an unbiased way bysetting constant thresholds and allowing Photoshop® to generate theimages that were quantitated to generate the data presented in FIG. 8.

These results show that there were statistically significant decreasesin infarct size when comparing the group pre-treated with normal waterto the groups pre-treated with water containing higher concentrations ofiodide (FIG. 8). These decreases were somewhat less or comparable tothose observed in mice intravenously administered iodide at the time ofreperfusion.

Example 5 Sodium Iodide Improves Chronic Heart Failure

This example demonstrates that treatment with sodium iodide (NaI)improves outcome in a mouse model of chronic heart failure.

In order to examine the effect of drinking water containing NaI onchronic heart failure following heart attack in a mouse model, twogroups of mice were provided with drinking water lacking iodide ordrinking water containing 0.77 mg/kg/day iodide starting immediatelyfollowing open chest ligation of the left descending coronary artery(LAD ligation) to initiate ischemia, and continuing until the end of thestudy, which lasted 12 weeks. Prior to the LAD ligation and at 1 week, 2weeks, 4 weeks, 8 weeks, and 12 weeks following the LAD ligation, micewere examined by echocardiography to determine their left ventricleejection fraction and their left ventricle fractional shortening. Theresults are shown in FIG. 9. This study was repeated as described abovewith more mice, and the combined results of both studies are shown inFIG. 10.

These results show that there were statistically significant increasesin left ventricle ejection fraction reduction and left ventriclefractional shortening in the group treated with water containing 0.77mg/kg/day iodide as compared to the group treated with normal drinkingwater (FIG. 9). These results demonstrate that treatment may be used totreat or prevent chronic heart failure, including chronic heart failurefollowing myocardial infarct.

Example 6 Sodium Iodide is Efficacious in the Treatment of AcuteMyocardial Infarction

This study demonstrates the efficacy of sodium iodide, administered at 1mg/kg as an i.v. bolus, in a rat model of acute myocardial infarction(AMI) by comparing 4 and 24 hr plasma cardiac troponin I (cTpnI) valuesand myocardial infarct sizes at 24 hrs between a placebo cohortreceiving saline only and subject to 30 min ischemia, an ischemicpreconditioned cohort (5 min ischemia/5 min reperfusion, 3×) subject to30 min of ischemia, and a treatment cohort receiving 1 mg/kg NaI as ani.v. bolus and subject to 30 min of ischemia.

Methods

Male CD® IGS Rats (300-450 g) were subjected to 30 minutes of LADocclusion followed by 24-30 hrs of reperfusion. 200-400 uL of blood wasdrawn at 4 and 24-30 hrs post-reperfusion for plasma cTpnI measurementsby EIA. After the 24-30 hr blood draw, the LAD was re-occluded, and theheart was excised following i.v. administration of Evans Blue. Thehearts were frozen overnight, sectioned, and stained with triphenyltetrazolium chloride for subsequent blinded infarct analysis.

Animal Preparation and Surgery

Rats were anesthetized with a ketamine/xylazine mixture (100 mg/kg and10 mg/kg, respectively) and maintained under 2-3% isoflurane in 100%medical oxygen after intubation with a 16 gauge angiocatheter sheath(Terumo® Surflo i.v. catheter, 16 G×2″, VWR Cat.# TESR-OX1651CA).Animals were ventilated using a Harvard Apparatus Model 683 small animalventilator equipped with a 5 cc cylinder at a tidal volume of 1.5-2.0mL/stroke and a respiratory rate of 105 strokes/min. With the rat lyingon its right side, animal fur was removed from the surgical area withclippers and then cleaned with alcohol pads. The heart beat wasvisualized through the chest wall and a dorsoventral incision in theleft chest wall was made directly over the beating heart. The underlyingfascia was dissected away to reveal the ribs and intercostal muscles. Aleft thoracotomy was then performed in the 4^(th) intercostal space andthe ribs were separated using alm retractors (Fine Science Tools Cat.#17008-07). The pericardium was then carefully opened and used to keepthe lungs and thymus out of the surgical field. The left anteriordescending coronary artery was visualized, and a 5-0 prolene suture withan RB-2 tapered needle (Ethicon Cat. #8710H) was passed underneath twiceat the boundary of the left atrial appendage to completely encircle theartery. The ends of the suture were passed through a 1 cm length ofPU-50 tubing, and ischemia was induced by tightening the suture againstthe tubing and securing in place with hemostats. Ischemia was verifiedby visual confirmation of cyanosis and akinesis in the left ventricularwall. Reperfusion was achieved by removing the hemostats and raising thetubing off of the ventricular wall, thereby loosening the suture aroundthe LAD. Reperfusion was confirmed by visualizing the reversal ofcyanosis in the ventricular wall. The suture around the LAD was left inplace, and the chest was then closed by re-approximating the edges ofthe ribs and stitching the musculature with a continuous 5-0 silksuture. As the last suture was being placed, the lungs were transientlyoverinflated by obstructing the expiratory limb of the respirator for3-4 cycles to expel air from the chest cavity. The suture was thentightened to seal the thoracic cavity, the facia over the ribs andintercostal muscles was sutured closed with 5-0 silk, and the skin wasthen sutured closed with 5-0 silk. The anesthesia was then turned off,and the animal was removed from the ventilator once spontaneousbreathing resumed. The endotracheal tube was removed once the animalregained consciousness. Over the course of the entire surgicalprocedure, the animals were on a heated, circulating water pad and undera heat lamp to maintain core body temperature. After recovery, allanimals remained in cages under heat lamps until the 4 hr blood draw, atwhich point they were placed back in the animal housing facility.

Plasma Sample Prep and cTpnI EIA

Under isoflurane anesthesia (3% for induction, 2% for maintenance), 4 hrblood draws were collected (by tail bleed for naïve and from carotidartery catheter for pre-cathed animals) into heparinized microtainerplasma separator tubes (VWR Cat. #VT365985) while 24-30 hr draws werecollected from either a carotid artery catheter (pre-cathed from CharlesRiver) or a jugular vein catheter (implanted in naïve animals prior toopening the chest wall and reoccluding the LAD). Blood was centrifugedat 10,000+ rpm for 5 min, and plasma was transferred to 0.5 mL microfugetube and stored at −20° C.

Cardiac troponin I was measured in a total of 51 plasma samples (26 4 hrsamples and 25 24-30 hr samples) which were run at 1:50 dilutions in aRat Cardiac Troponin-I, High Sensitivity ELISA (Kamiya BiomedicalCompany Cat. #KT-639) according to the manufacturer's instructions.

Morphometric Analysis

After 24 hr blood draws, the chest wall was reopened through theprevious day's incision, and the LAD was re-occluded using the prolenesuture left in place around the LAD the prior day. Once ischemia wasobserved visually, a 10% Evans Blue solution was administered i.v. (viacarotid or jugular cannula) until the perfused area of the heart turnedblue, and then the heart was excised from the thoracic cavity. Atria,valves, large vessels, and right ventricle were trimmed away afterthorough rinsing in water, and the left ventricle was then rinsedbriefly again with water, wrapped in cellophane, and frozen overnight at−20° C. The left ventricle was then sliced into six 2 mm thick slicesand incubated in 1.5 mL 1% TTC in PBS for 20 min. at 37° C. Images ofthe apical and basal surfaces of each slice were then obtained on aNikon DSL 3200 equipped with a macro lens. Infarct analysis was blindedwith respect to treatment.

Results

Plasma troponin levels were consistently lower in animals treated withsodium iodide than those treated with placebo (FIG. 10). In addition,infarcts were smaller in animals treated with sodium iodide as comparedto those treated with placebo (FIG. 11), whereas infarcts in theischemic preconditioning cohort showed onlyu nonsignificant trends inreduced size as compared to placebo. These data demonstrate thatadministration of iodide is effective in treating AMI.

Example 7 Treatment of Human Disease with Elemental Reducing Agents

A variety of human diseases are associated with the formation ofreactive oxygen species that cause lipid peroxidation and proteincarbonylation, including those diseases shown in the previous examplesto benefit from treatment with iodide (or sulfide or selenide). Thesechemical modifications of biopolymers play a causal role in the loss ofcells resulting in disease and death. Accordingly, other diseasesassociated with the formation of reactive oxygen species may also betreated using iodide or another elemental reducing agent (ERA),including any of those described herein, e.g., sulfide or selenide. Thefollowing diseases are tested in animal or ex vivo models of humandisease to demonstrate the outcomes expected when the animals or tissuesare treated with iodide or another ERA such as sulfide or selenide. Allstudies include intravenous treatment with ERAs.

Renal Transplant

Kidneys are removed from dogs and pigs, stored for 24 hours and thentransplanted into an animal in which they lack a second kidney and,therefore, are dependent on the function of the transplanted kidney.ERAs, e.g., iodide, sulfide, and selenide, are expected to increase thefunctionality of the stored kidney, establishing that ERAs can be usedto improve human kidney transplantation.

Graft Vs. Host Disease

Mice and dogs are irradiated to destroy bone marrow stem cells (BMSC),and unmatched donor BMSC are transplanted into the irradiated animals inthe absence of cyclosporine A treatment. It is expected that ERAs, e.g.,iodide, sulfide and selenide, will function as cyclosporine and enableunrelated marrow to engraft without graft rejection and loss of life.

Peripheral Vascular Disease

Mice are surgerized to prevent femoral blood flow to the hind limb.Treatment with an ERA, e.g., iodide, sulfide or selenide, twice dailyfor 2 weeks following surgery is expected to permanently improve bloodflow.

Noise-Induced Hearing Loss

Mice are exposed to loud noise and then intravenously treated with anERA, such as iodide, selenide or sulfide. It is expected that treatmentwith the ERA will improve outcome compared to untreated control animals,i.e., resulting in decreased hearing loss.

Acute Ischemic Stroke

Rats exposed to temporary loss of blood flow to the brain are reperfusedat the same time they are treated with an ERA, such as iodide, sulfideor selenide. It is expected that treatment with the ERA will reducetissue damage caused by the loss of blood flow, and reduceischemic-reperfusion infury.

Acute Myocardial Infarction

The left anterior descending coronary artery of animals (mice, rats andrabbits) is ligated temporarily to mimic a heart attack. At the point ofreperfusion, an ERA such as iodide, sulfide or selenide is infused. Thistreatment decreases heart damage and improves heart function.

Contrast-Induced Nephropathy

Mice are treated with a contrast agent under conditions that inducekidney injury to model the type of injury frequently suffered bypatients that are exposed to an intravenous contrast agent in the courseof vascular imaging. It is expected that treatment with an ERA, e.g.,iodide, sulfide or selenide, will reduce damage and improve kidneyfunction.

Sickle Cell Anemia

Mice bearing specific mutant hemoglobin suffer similar red cellhemolysis and vascular damage found in patients with sickle cell anemia.Damage in these mice is exacerbated by acute hypoxic exposure. Thesemice will be treated with an ERA such as sulfide, selenide or iodide inthe presence or absence of acute hypoxic exposure, and it is expectedthat treatment with the ERA will protect them from this damage.

Postoperative Ileus

Pertioneal surgery is associated with extended recovery owing to aperiod in which there is a lack of intestinal peristalsis. Such a lackof peristalsis is induced in mice, and the mice are then treated with anERA, e.g., iodide, sulfide or selenide. It is expected that the ERA willshorten/eliminate this lack of peristalsis following surgery.

Radiation Induced Lung Injury

Radiation treatment in the course of cancer therapy sometimes results inunintended lung injury. Mice are irradiated to generate lung injury. AnERA, e.g., iodide, sulfide or selenide, is administered to the miceprior to and following irradiation. It is expected that treatment withthe ERA will decrease lung injury.

Ventilator Induced Lung Injury

Lung ventilation is of great value for some patients; however,ventilation can damage the lung. Mice are exposed to ventilation injurywith or without treatment with an ERA such as iodide, sulfide orselenide. It is expected that ERA treatment will decrease lung injury.

Salmonella Infection

Bacterial infection with Salmonella can be lethal. Mice are infectedwith Salmonella and treated with an ERA, e.g., iodide, sulfide orselenide. It is expected that the ERA will suppress immune andinflammatory responses, thereby improving survival.

Retinopathy

Loss of cells in the retina is thought to come about from reactiveoxygen species. ERAs are anti-oxidants. Rats are treated with conditionsthat cause retinopathy modeling human retinal degeneration and treatedwith an ERA such as iodide, sulfide or selenide. It is expected thattreatment with the ERA will improve retinal cell viability

Organ Preservation

Organ transplantation involves temporary loss of blood flow. Explantedorgans from donor pigs are perfused ex vivo with an ERA, e.g., iodide,sulfide or selenide, in preservation solution before being transplantedinto a recipient pig. It is expected that treatment with the ERA willimprove the functioning of the organ once it is transplanted into therecipient.

Hypoxia-Ischemia Encephalopathy

Temporary loss of blood flow, followed by reperfusion can result inpermanent brain damage. Newborn ferrets are treated with an ERA, e.g.,iodide, sulfide or selenide, under conditions that mimic this braindamage. It is expected that treatment with the ERA at birth willdecrease brain damage.

Epilepsy

Epilepsy is associated with seizures causing excessive metabolicactivity in the brain and damage. ERA treatment decreases metabolicrate. Rats are treated with an ERA, e.g., iodide, sulfide or selenide,under conditions that model epileptic seizures and brain damage. It isexpected that ERA treatment of rats suffering seizures will decreasebrain damage.

Stem Cell Engraftment

Transplantation of stem cells results in oxidative damage and loss ofmany of the cells. Cardiomyocytes are transplanted into damaged rathearts, and the cells and animals are either treated with an ERA, e.g.,iodide, sulfide or sulfide, or left untreated. It is expected thattreatment of the stem cells and the recipient animals with an ERA willimprove stem cell engraftment.

All of the above U.S. patents, U.S. patent application publications,U.S. patent applications, foreign patents, foreign patent applicationsand non-patent publications referred to in this specification and/orlisted in the Application Data Sheet, are incorporated herein byreference, in their entirety.

We claim:
 1. A method for treating or inhibiting a reperfusion injuryresulting from restoration of blood flow to an ischemic tissue or organin a subject, comprising providing to said subject a pharmaceuticalcomposition comprising sodium iodide and a pharmaceutically acceptablecarrier, diluent, or excipient, wherein said composition is provided tothe subject in an amount sufficient to increase the blood concentrationof iodide in the subject by at least 500% for at least some time.
 2. Themethod of claim 1, wherein said composition is provided to the subjectparenterally.
 3. The method of claim 2, wherein said composition isprovided to the subject intravenously.
 4. The method of claim 1, whereinsaid reperfusion injury occurs following percutaneous coronaryintervention or thrombolysis.
 5. The method of claim 1, wherein thesubject is provided with the pharmaceutical composition daily, twicedaily, or once every two days.
 6. The method of claim 5, wherein thesubject is provided with a dose of about 0.1 mg/kg to about 5 mg/kg ofthe sodium iodide.
 7. The method of claim 6, wherein the subject isprovided with a dose of about 0.5 mg/kg to about 2 mg/kg of the sodiumiodide.
 8. The method of claim 7, wherein the subject is provided with adose of about 1.0 mg/kg of the sodium iodide.